Therapeutic systems, devices, and compositions with wound healing and tissue regenerative properties, uses thereof, and corresponding methods

ABSTRACT

Presented herein are compositions that can be administered to a subject having damaged tissue, for example a wound. The compositions are often administered in combination with administration of energy (e.g., laser energy, light from a light emitting diode, radiofrequency (RF) energy, audio frequency energy, etc.) from an energy generating device and/or system to the affected site. The compositions, systems, devices, and methods herein were found to induce wound healing and tissue regeneration.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to U.S. provisional patent application62/885,761, filed Aug. 12, 2019, and U.S. provisional patent application62/992,579, filed Mar. 20, 2020, the entire contents of each of whichare herein incorporated by reference in their entireties.

FIELD OF THE INVENTION

Therapeutic systems, devices, compositions, and methods are describedthat can be used for the treatment of damaged tissue, which methodsinclude, in some embodiments, administration of a therapeuticcomposition, with or without application of additional energy (e.g.,laser energy, light from a light emitting diode, radiofrequency (RF)energy, audio frequency energy, etc.), to a damaged tissue.

BACKGROUND

Wound healing systems are known. Typically such systems deliver highenergy radiation to tissue to weld the tissue together and/or provide anadhesive configured to join different portions of skin together.

SUMMARY

In certain aspects, provided herein are compositions comprising two ormore of: collagen, hyaluronic acid, fucose, copper, and iron. In someembodiments, the compositions here are used to treat wounds or tissuedamage.

In some aspects, provided herein are systems for treating wounds ortissue damage. In some embodiments, the systems comprise one or moreenergy sources and/or devices, one or more of the compositions describedherein, an imaging device, and/or other components.

In some aspects, provided herein are methods of treating wounds ortissue damage. In some embodiments, the methods comprise administering atherapeutically effective amount of a composition described herein tothe wound or damaged tissue and/or administering a therapeuticallyeffective amount of energy (e.g., laser energy, light from a lightemitting diode, radiofrequency (RF) energy, audio frequency energy,etc.) to the wound or damaged tissue.

In some aspects, provided herein are devices configured to generate oneor more types of therapeutic energy for provision to tissue. In someembodiments, the devices comprise one or more energy sources configuredto generate one or more types of therapeutic energy. The one or moretypes of therapeutic energy may comprise one or more of laser radiation,radio frequency (RF) waves, audio frequency waves, or light from a lightemitting diode (LED).

In some aspects, provided herein are devices for imaging tissue. In someembodiments, the devices comprise one or more light emitting diodes(LED) configured to emit light for illuminating and reflecting thetissue, a camera configured to obtain images of the illuminated tissue,one or more processors configured to analyze the images, and/or othercomponents.

BRIEF DESCRIPTION OF THE DRAWINGS

The drawings illustrate embodiments of the technology and are notlimiting. For clarity and ease of illustration, the drawings are notmade to scale and, in some instances, various aspects may be shownexaggerated or enlarged to facilitate an understanding of particularembodiments.

FIG. 1 shows bone profile changes for a zygomatic arch treated with acomposition comprising hyaluronic acid, iron, and copper gluconate.Cephalometric x-rays were overlaid at different points along thezygomatic arch (E, F, G, H and I) were measured.

FIG. 2 shows bone profile changes of a facial zygomatic arch before andafter treatment with a composition comprising hyaluronic acid, iron, andcopper gluconate. Cephalometric x-rays were overlaid at different pointsalong the zygomatic arch (A, B, C, D) were measured.

FIG. 3 shows bone profile changes for a zygomatic arch treated with acomposition comprising hyaluronic acid, iron, and copper gluconate.Cephalometric x-rays were overlaid at different points along thezygomatic arch (E, F, G, H and I) were measured. FIG. 4 shows boneprofile changes.

FIG. 4 shows bone profile changes of a facial zygomatic arch before andafter treatment with a composition comprising hyaluronic acid, iron, andcopper gluconate. Cephalometric x-rays were overlaid at different pointsalong the zygomatic arch (A, B, C, D) were measured.

FIG. 5 shows tissue regeneration differences before (left bar of eachpair) and after (right bar of each pair) treatment of maxillary anteriorgingival with a composition comprising hyaluronic acid, iron, and coppergluconate. Before and after pictures were taken and anterior attachedgingiva was measured along 4 points (before and after).

FIG. 6 shows changes in epithelium crease length before and aftertreatment with a composition comprising hyaluronic acid, iron, andcopper gluconate. Pictures were taken of a Leg keloid scar before andafter treatment and scar creases were measured along 6 points (beforeand after).

FIG. 7 shows changes in epithelium crease length before (left bar ofeach pair) and after (right bar of each pair) treatment with acomposition comprising hyaluronic acid, iron, and copper gluconate.Pictures were taken of a Leg keloid scar before and after treatment andscar creases were measured along 5 points (before and after).

FIG. 8 shows changes in epithelium crease depth before (left bar of eachpair) and after (right bar of each pair) treatment with a compositioncomprising hyaluronic acid, iron, and copper gluconate. Pictures weretaken of a Leg keloid scar before and after treatment and scar creasedepth was measured along 7 points (before and after).

FIG. 9 shows soft tissue repair of a facial scar and shows changes inepithelium crease depth before (left bar of each pair) and after (rightbar of each pair) treatment with a composition comprising hyaluronicacid, iron, and copper gluconate. Pictures were taken of the facial scarbefore and after treatment and scar depths were measured along 3 points(before and after).

FIG. 10 shows subjective data for mucosal before and after treatmentwith a composition comprising hyaluronic acid, iron, and coppergluconate. Patients (P1 and P2) were given a questionnaire before (leftthree bars of each set) and after (right three bars of each set)treatment. The questions related to dryness (bars 1 and 4 of each set,as count from left to right), comfort (bars 2 and 5 of each set, ascount from left to right) and pain (bars 3 and 6 of each set, as countfrom left to right).

FIG. 11 shows changes in epithelium crease length and depth of a buttockscar before and after treatment with a composition comprising hyaluronicacid, iron, and copper gluconate. Pictures were taken of the buttockscar before and after treatment, and scar crease length and depth weremeasured along 4 points (before and after).

FIG. 12 shows changes in epithelium crease length of an arm scar before(bottom bar of each pair) and after (top bar of each pair) treatmentwith a composition comprising hyaluronic acid, iron, and coppergluconate. Pictures were taken of the arm scar before and aftertreatment, and scar crease length was measured along 2 points (beforeand after).

FIG. 13 shows changes in epithelium crease length of a leg scar before(bottom bar of each pair) and after (top bar of each pair) treatmentwith a composition comprising hyaluronic acid, iron, and coppergluconate. Pictures were taken of the leg scar before and aftertreatment, and scar crease length was measured along 7 points (beforeand after).

FIG. 14 shows changes in arm pit firmness before (top bar of each pair)and after (bottom bar of each pair) treatment with a compositioncomprising hyaluronic acid, iron, and copper gluconate. Armpit firmnesswas assessed by a subjective questionnaire provided before and aftertreatment for the two data points reported.

FIG. 15 shows changes in scar area (first bar of each set of three),length (second bar of each set of three) and width (third bar of eachset of three) of a stomach scar before (left set of three bars) andafter (right set of three bars) treatment with a composition comprisinghyaluronic acid, iron, and copper gluconate. Pictures were taken of thestomach scar before and after treatment.

FIG. 16 shows changes in epithelium crease length of a leg keloid scarbefore (left bar of each pair) and after (right bar of each pair)treatment with a composition comprising hyaluronic acid, iron, andcopper gluconate. Pictures were taken of the leg scar before and aftertreatment, and scar crease length was measured along 7 points (beforeand after).

FIG. 17 shows changes in an open leg wound depth before (top bar of eachpair) and after (bottom bar of each pair) treatment with a compositioncomprising hyaluronic acid, iron, and copper gluconate. Pictures weretaken of the leg wound before and after treatment and wound depth wasmeasured along 4 points (F, G, H and I, before and after).

FIG. 18 shows changes in an open leg wound depth before (right backbars) and after (left forward bars) treatment with a compositioncomprising hyaluronic acid, iron, and copper gluconate. Pictures weretaken of the leg wound before and after treatment and wound depth wasmeasured along 4 points (F, G, H and I, before and after).

FIG. 19 shows changes in a leg scar length before (bottom bar) and after(top bar) treatment with a composition comprising hyaluronic acid, iron,and copper gluconate. Pictures were taken of the leg scar before andafter treatment and scar length was measured along 2 points (before andafter).

FIG. 20 A-D shows bone densities (bone growth) after treatment with acomposition comprising collagen, iron, and copper gluconate. Allpercentages are percentages of opaqueness on an x-ray. All percentagesof opaqueness were measured by measuring the same points along eachx-ray each month. When bone grows, the dark x-ray turns to opaque.Different dark points measured (no bone present) turned opaque (bonegrowth) with measured time.

FIG. 21 is an electrical drawing of a possible example of a functiongenerator which generates audio and/or radiofrequency Hz signals.

FIG. 22 is a mechanical drawing of a possible example of a functiongenerator which generates audio and/or radiofrequency signals.

FIG. 23 illustrates a possible example of an LED board that includes aplurality of LEDs that can be used to treat and/or image tissue.

FIG. 24a-24c show various views of an example embodiment of a deviceconfigured to generate radiofrequency energy; FIG. 24a shows a frontperspective view. FIG. 24b shows a right side view. FIG. 24c shows abottom view.

FIG. 25a-25d show views of the example embodiment of the RF or audiodevice 200 with and without a laser energy source. FIG. 25a shows anexploded view of the RF or audio device with a laser. FIG. 25b shows aclose-up of a housing (e.g., a handpiece) with a laser relative to thehousing. FIG. 25c shows an exploded view of an RF or audio devicewithout a laser. FIG. 25d shows a detailed view of a housing for an RFor audio device without a laser.

FIG. 26 illustrates a flow chart for an example method for using an RFor audio device.

FIGS. 27a-27e show various views of another example embodiment of an RFor audio device. FIG. 27a shows a top view. FIG. 27b shows a side view.FIG. 27c shows a perspective view of the RF or audio tips. FIG. 27dshows an exploded side perspective view. FIG. 27e shows an alternativeside perspective view.

FIG. 28 is an electrical drawing of a possible example of circuitry thatmay be included in a device described herein.

FIG. 29 is a schematic illustration of an example of a possible devicethat uses laser energy for treatment of tissue.

FIG. 30 illustrates an example of laser fibers (e.g., conduits) that maybe included in a device that uses laser energy for treatment of tissue.

FIG. 31 illustrates combining individual laser fibers for a device thatuses laser energy for treatment of tissue.

FIG. 32 illustrates an example of components of a possible embodiment ofan LED device that may be included in a present system and/or used in apresent method.

FIG. 33 illustrates another example of components of a possibleembodiment of an LED device that may be included in a present systemand/or used in a present method.

FIG. 34 illustrates an example of components of an imaging device thatmay be included in a present system and/or used in a present method.

FIG. 35 shows a table of results for the treatment of tooth extractionsites with the substrates (compositions) of formulas 7, 12, 16-18, 22and 23 alone (without energy (e.g., laser energy, light from a lightemitting diode, radiofrequency (RF) energy, audio frequency energy,etc.)).

FIG. 36 shows a table of results for the treatment of tooth extractionsites with the substrates (compositions) of formulas 7, 12, 16-18, 22and 23 and treatment with energy (e.g., laser energy, light from a lightemitting diode, radiofrequency (RF) energy, audio frequency energy,etc.).

FIG. 37 shows a table of results for the treatment of leg wounds withthe substrates (compositions) of formulas 7, 12, 16-18, 22 and 23 alone(without energy (e.g., laser energy, light from a light emitting diode,radiofrequency (RF) energy, audio frequency energy, etc.)).

FIG. 38 shows a table of results for the treatment of leg wounds withthe substrates (compositions) of formulas 7, 12, 16-18, 22 and 23 andtreatment with energy (e.g., laser energy, light from a light emittingdiode, radiofrequency (RF) energy, audio frequency energy, etc.).

FIG. 39 shows a table of results for the treatment of tooth extractionsites (Extraction Site) and leg wounds with energy (e.g., laser energy,light from a light emitting diode, radiofrequency (RF) energy, audiofrequency energy, etc.) in the absence of a composition (e.g., asubstrate).

FIG. 40 shows a table of results for the treatment of acute wounds. Oralsurgery acute wounds are demarked by very small changes (e.g., 4, 5, 6,7, 14, 22 and 23 mm) and all others are leg chronic open wounds demarkedby large mm changes with the substrates (compositions) of formulas 1-23and treatment with energy. For oral surgery: extraction/s wereperformed, a laser-RF or audio device (described herein) was placed overthe extraction site, the unit was turned on, followed by emissionthrough the device for 1-2 minutes. A substrate (composition) was placedinto the extraction site. The laser-RF or audio device was placed overthe extraction site, turned on, and emission through the device beganfor 1-2 minutes. The patient was allowed to keep their mouth in an openposition for 2-5 minutes until clear carbon begins to form. For achronic wound—substrate was placed onto the wound. Laser-rf wounds withemission from handpiece for 15 minutes. Optionally repeat several timesuntil the surrounding tissue around the wound and the surface of thewound begins to firm. Wound tape was placed onto the wound. An absorbentpad was placed over the taped wound. The wound site and tissue wasimmobilized with sports tape.

FIG. 41 shows a table of results for the treatment of acute wounds withthe substrates of formulas 1-23 alone (without energy (e.g., laserenergy, light from a light emitting diode, radiofrequency (RF) energy,audio frequency energy, etc.)). In this table, the row of formulation 3is a head wound; the row of formulation 4 is a leg wound; the rows offormulations 5, 6 and 15-18 are extractions; and the rows formulas 13and 19-21 are oral surgery sites in the mucosa.

FIG. 42 shows a table of results for the treatment of chronic woundswith the substrates of formulas 1-23 alone (without energy (e.g., laserenergy, light from a light emitting diode, radiofrequency (RF) energy,audio frequency energy, etc.)). In this table, the rows of formulas 1,4, 5, 7 and 8 are periodontal wounds. The remaining rows are leg wounds.

FIG. 43 shows a table of results for the treatment of chronic woundscomprising chronic open leg wounds with the substrates of formulas 1-23and treatment with energy (e.g., laser energy, light from a lightemitting diode, radiofrequency (RF) energy, audio frequency energy,etc.). In this example, the energy comprises laser energy. To generatethe data in FIG. 43, a substrate (composition) was placed onto wound.Laser energy was applied to the wound with laser emission from a laserdevice for 15 minutes. This was optionally repeated several times untilthe surrounding tissue around the wound and the surface of the woundbegan to firm. Wound tape was placed onto the wound. An absorbent padwas placed over the taped wound. The wound site was immobilized withsports tape.

FIG. 44 shows a table of results for the treatment of chronic wounds.

FIG. 45 shows a table of results for the treatment of a chronic woundcomprising a chronic leg wound with RF or audio in the absence of asubstrate (composition). To generate the data in the table of FIG. 45,an RF or audio device was placed over the wound with emission from thedevice for 15 minutes. This was optionally repeated several times untilthe surrounding tissue around the wound and the surface of wound beganto firm. Wound tape was placed onto the wound. An absorbent pad wasplaced over the taped wound. The wound site and surrounding tissue wasimmobilized with sports tape.

FIG. 46 shows a table of results for the treatment of a chronic woundsuch as a chronic leg wound with the substrate (composition) of formula21 and RF or audio. To generate the data for the table of FIG. 46, an RFor audio device was placed over the wound with emission from the devicefor 15 minutes. This was optionally repeated several times until thesurrounding tissue around the wound and the surface of wound began tofirm. Wound tape was placed onto the wound. An absorbent pad was placedover the taped wound. The wound site and surrounding tissue wasimmobilized with sports tape.

FIG. 47 shows a table of the composition of the substrates of formula1-23, formulated as a liquid, to be used for prophylactic supplement.

The formulations (compositions) used to generate the data of FIGS. 35-46were formulated as a gel.

DETAILED DESCRIPTION

Provided herein are systems, compositions, devices, and methods for thetreatment of tissue damage and/or to induce tissue regeneration.Compositions disclosed herein may comprise two or more of collagen,hyaluronic acid, fucose, copper, iron, silver, gold, ATP andacetylcholine. In some embodiments, a composition comprises three ormore ingredients selected from collagen, hyaluronic acid, fucose,copper, iron, silver, gold, ATP and acetylcholine. In some embodiments,a composition comprises four, five, six, seven, or eight ingredientsselected from collagen, hyaluronic acid, fucose, copper, iron, silver,gold, ATP and acetylcholine. In certain embodiments, a compositioncomprises collagen, hyaluronic acid, fucose, copper, iron, silver, gold,ATP and acetylcholine. A system and/or a device may be and/or formdelivery means of one or more substrates (and/or one or more portions ofa substrate (e.g., the collagen or Hyaluronic acid) to a wound site. Agiven system and/or device may be configured such that one or morecompositions (and/or portions of compositions) are delivered to cells bymeans of the energy generated by the device and/or system, and/or byother means.

In some embodiments, a composition comprises collagen and one, two,three, four or more ingredients selected from hyaluronic acid, fucose,copper, iron, silver, gold, ATP and acetylcholine. In some embodiments,a composition comprises hyaluronic acid and one, two, three, four ormore ingredients selected from collagen, fucose, copper, iron, silver,gold, ATP and acetylcholine. In some embodiments, a compositioncomprises hyaluronic acid and one, two, three, four or more ingredientsselected from collagen, fucose, copper, iron, silver, gold, ATP andacetylcholine.

Compositions disclosed herein can be used alone or in combination withadministration of energy (e.g., laser energy, light from a lightemitting diode, radiofrequency (RF) energy, audio frequency energy,etc.) to a damaged tissue to induce wound healing and/or tissueregeneration. Compositions disclosed herein are sometimes referred toherein as “substrates”. One or more compositions or substrates asdescribed herein may be included as a component in a system that alsoincludes an energy source or device, and/or other components.

Subjects

The term “subject” refers to a mammal. Any suitable mammal can betreated by a method or composition described herein. Non-limitingexamples of mammals include a human, non-human primate (e.g., ape,gibbons, chimpanzees, orangutans, monkeys, macaques, and the like),domestic animals (e.g., dogs and cats), farm animals (e.g., horses,cows, goats, sheep, pigs) and experimental animals (e.g., mouse, rat,rabbit, guinea pig). In some embodiments a subject is a non-humanprimate or a human. In some embodiments a subject is a human. A subjectcan be any age or at any stage of development (e.g., an adult, teen,child, infant, or a mammal in utero). A subject can be male or female.

Damaged Tissue

In some embodiments, a composition or method herein is used to treatdamaged tissue. In some embodiments, a damaged tissue is a wound,non-limiting examples of which include a laceration, cut, scrape,abrasion, scratch, puncture, bite (e.g., an insect or animal bite), burn(e.g., a chemical burn, temperature-induced burn (e.g., frost bite, sunburn, a burn by heat or fire)), bruise or contusion, ulcer, cyst,sprain, muscle tear or pull, ligament tear or pull, a bone fracture orbreak, a surgical incision site, a surgical cut site, a surgicalextraction site. In some embodiment, a wound is a tooth extraction site.In certain embodiments, a wound is an acute wound. In certainembodiments, an acute wound is a wet wound. In certain embodiments, awound is a chronic wound. In certain embodiments, a chronic wound is adry wound. In some embodiments, a damaged tissue is hypoxic. In someembodiments, a damaged tissue is ischemic, lacks sufficient circulation,and/or is restricted of blood flow. In some embodiments, damaged tissuecomprises tissue lacking collagen integrity, structure or form. In someembodiments, damaged tissue comprises tissue that has lost electrons andtherefore damaged due to aging. In some embodiments, a damaged tissuecomprises a vasculature obstruction.

In some embodiments, a damaged tissue is a necrotic tissue. In someembodiments, a damaged tissue is a scar. A damaged tissue may becancerous, diseased (e.g., gum disease), aged, aging, damaged, necrotic,infected, mature, immature, normal, abnormal, missing, underdeveloped,developed, clean, dirty, or contaminated. In certain embodiments, awound is any area of a subject that has lost any amount of originaltissue or bone.

In certain embodiments, a wound is an ulcer. In some embodiments, awound is a diabetic ulcer. In some embodiments, a wound is a non-healingwound, for example a wound on a diabetic subject that is not healing(e.g., a chronic burn wound, or chronic skin ulcer). In someembodiments, a wound is a venous ulcer.

A damaged tissue may be of any size. In some embodiment, a wound has amean, median, average or absolute diameter of 0.1 mm to 60 cm or more.In some embodiment, a wound has a mean, median, average or absolutediameter of 0.1 mm to 25 mm, 0.1 mm to 15 mm, 0.1 mm to 10 mm, or 0.1 mmto 5 mm. In some embodiment, a wound has a mean, median, average orabsolute depth of 0.1 mm to 25 mm, 0.1 mm to 15 mm, 0.1 mm to 10 mm, or0.1 mm to 5 mm.

A damaged tissue may be of any age. In some embodiment, a wound is new,for example a wound that appears within 1 day, 1-4 days or up to a week.In some embodiment, a wound is about 4 or more, 6 or more, 20 or more or60 or more days old. In some embodiment, a wound is new, for example awound that appears within 1 day, 1-4 days or up to a week.

In some embodiments, a chronic wound is an ulcer or bed sore. In someembodiments, a chronic wound is a diabetic ulcer.

In some embodiments, a damaged tissue comprises endothelium, epithelialtissue, connective tissue (e.g., cartilage or bone), muscle tissue(e.g., smooth muscle, skeletal muscle or cardiac muscle tissue), nervoustissue, and/or combinations thereof. In some embodiments, a damagedtissue comprises damaged bone. In some embodiments, a damaged tissuecomprises damaged epithelial tissue. In some embodiments, a damagedtissue comprises damaged veins or arteries.

Energy

In some embodiments, a damaged tissue is treated with energy (e.g.,laser energy, light from a light emitting diode, radiofrequency (RF)energy, audio frequency energy, etc.). The energy may be and/or includeelectromagnetic radiation and/or other energy. The radiation maycomprise and/or be associated with photons, electrons, audio waves,radio frequency waves, magnetic waves, electricity, and/or other energy.In some embodiments, a damaged tissue is treated with electromagneticradiation and one or more compositions described herein. Electromagneticradiation may be produced by one or more suitable energy sources ordevices, non-limiting examples of which include a lasers, a laser photonbeam, lamps, a bulb, a tube, a diode, a light emitting diode (LED), LEDphoton beam, broadband photon beam, radiofrequency electron waveforms,audio frequency waveforms, electrical discharge, the like, andcombinations thereof.

In some embodiments, electromagnetic radiation that is used for a systemand/or method described herein is at a wavelength and/or frequency inthe visible spectrum. In some embodiments, electromagnetic radiationthat is used for a system and/or a method described herein is at awavelength and/or frequency in the infrared spectrum. In someembodiments, electromagnetic radiation that is used for a system and/ora method described herein is at a wavelength and/or frequency in theultraviolet spectrum. In some embodiments, electromagnetic radiationthat is used for a system and/or a method described herein is at awavelength and frequency in the radio frequency (RF) spectrum ofelectromagnetic radiation. In some embodiments, the energy that is usedfor a system and/or a method described herein is at a wavelength andfrequency in the audio frequency spectrum. In some embodiments,electromagnetic radiation that is used for a system and/or a methoddescribed herein is at one or more wavelengths, frequencies, and/orpowers described herein, and combinations thereof.

In some embodiments, energy (e.g., laser energy, light from a lightemitting diode, radiofrequency (RF) energy, audio frequency energy,etc.) that is used for a system and/or a method described hereincomprises a wavelength between about 10 nm and 100 nm, or intermediateranges therein. In some embodiments, energy (e.g., laser energy, lightfrom a light emitting diode, radiofrequency (RF) energy, audio frequencyenergy, etc.) that is used for a system and/or a method described hereincomprises a wavelength between about 10 nm and 100 nm, or intermediateranges therein. In some embodiments, energy (e.g., laser energy, lightfrom a light emitting diode, radiofrequency (RF) energy, audio frequencyenergy, etc.) that is used for a system and/or a method described hereincomprises a wavelength between about 10 nm and 1800 nm, between about100 nm and 1800 nm, between about 200 nm and 1200 nm, between about 200nm and about 2500 nm, between about 380 nm and 1200 nm, between about400 nm and 1200 nm or between about 450 nm and 1200 nm. In someembodiments, energy (e.g., laser energy, light from a light emittingdiode, radiofrequency (RF) energy, audio frequency energy, etc.) that isused for a system and/or a method described herein comprises awavelength between about 600-1200 nm, 700-1400 nm, 700-1200 nm, 750-1000nm, 800-900 nm, 520-570 nm, 620-750 nm or 570-590 nm. In someembodiments, energy (e.g., laser energy, light from a light emittingdiode, radiofrequency (RF) energy, audio frequency energy, etc.) that isused for a system and/or a method described herein comprises awavelength between about 200 nm and 2500 nm. In some embodiments, energy(e.g., laser energy, light from a light emitting diode, radiofrequency(RF) energy, audio frequency energy, etc.) that is used for a systemand/or a method described herein comprise about a 1 Hz to about a 2.4GHz frequency range.

In some embodiments, energy that is used for a system and/or a methoddescribed herein comprises a wavelength in the radio frequency (RF)range, for example between about 1 Hz to about 2.4 GHz. In someembodiments, RF frequency that is used for a method herein comprises awavelength in a range of about 1 Hz to about 2.4 GHz.

In some embodiments, energy that is used for a system and/or a methoddescribed herein provides an energy (i.e., a dose) to a region of tissuedamage of about 1 Joule/cm² to about 400 Joule/cm², about 1 Joule/cm² toabout 150 Joule/cm², about 1 Joule/cm² to about 100 Joule/cm², about 10Joule/cm² to about 150 Joule/cm², or about 20 Joule/cm² to about 100Joule/cm².

In some embodiments, a laser or light source providing electromagneticradiation (e.g., energy) to a region of tissue damage comprises a powerof about 0.0001 W to about 100 W, 0.0001 to about 40 W, about 0.0001 toabout 10 W, about 0.0001 to about 5 W, or about 0.001 to about 1 W perwaveform, beam or current. In some embodiments, an infrared light sourcecomprises a power of about 0.001 W-5 W. In some embodiments, an RFsource comprises a power of about 0.001 W to about 10 W. In someembodiments, an RF source comprises a power of about 10 W or less, 9 Wor less, 8 W or less, 6 W or less, or 5 W or less. In some embodiments,an RF source emits at a frequency in a range of 3 Hz to 30 GHz, 0.5 MHzto 24 GHz, 0.1 MHz to 40 MHz, 0.1 MHz to 20 MHz, 0.2-10 MHz, 0.3-5 MHz,or 0.5 KHz to 40 KHz. In some embodiments, an RF source emits at afrequency in a range of 500 KHz or less.

Devices

A therapeutically effective amount of energy (e.g., laser energy, lightfrom a light emitting diode, radiofrequency (RF) energy, audio frequencyenergy, etc.) can be applied to a damaged tissue using a suitabledevice. A device may include an energy source configured to produce theenergy described here, electronic circuitry, a housing, one or morecomponents configured to direct the energy, and/or other components. Insome embodiments, a device that can be used in a system and/or for amethod described herein is shown in one or more of FIGS. 21-34. In someembodiments, example devices (e.g., laser, RF, LED, audio frequency,etc.), and/or device components, similar to and/or the same as thedevices and/or device components described herein that can be used in asystem and/or for a method described herein are described in U.S. patentapplication Ser. No. 14/937,858 (titled “Laser Assisted Wound HealingProtocol and System” and filed on Nov. 10, 2015); U.S. patentapplication Ser. No. 15/348,793 (titled “Laser Assisted Wound HealingProtocol and System” and filed on Nov. 10, 2016); and/or U.S. patentapplication Ser. No. 15/811,651 (titled “Laser Assisted Wound HealingProtocol and System” and filed on Nov. 13, 2017); which are eachincorporated by reference in their entirety herein.

In some embodiments a device comprises an energy source capable ofemitting a therapeutically effective amount of energy to a damagedtissue. In some embodiments, a device comprises more than one energysource capable of emitting different types of energy (e.g., laser, RF,audio frequency, LED energy, etc.), energy having different wavelengths(e.g., energy comprising dual wavelengths may be produced by a singledevice), different powers, different doses, and/or othercharacteristics. In some embodiments, a device comprises one or more ofa handpiece (e.g., a housing), an energy source (e.g., laser, LED, lamp,bulb, an RF source, an audio frequency source (e.g., a speaker), etc.),an aperture, one or more lenses, fiber optics, a user interface, acontrol panel, a control pad, a conduit, and/or other components. Insome embodiments, a device may include two or more different energysources. For example, a device may include an LED, a laser energysource, an RF source, an audio frequency source, some combination of twoof those, a combination of any three, or a combination of all four. Insome embodiments, a device may include and/or produce one or moreindividual beams and/or waves (e.g., the terms “beam” and “wave” are notintended to be limiting) of energy of one or more of the different typesof energy described herein. These beams and/or waves may be output by adevice through one or more fibers, openings in a device housing (e.g.,handpiece), and/or by other means.

By way of several non-limiting examples, in some embodiments, a devicemay have one or more of an energy source, an opening (e.g., forgenerated energy to leave the device), a hand piece, a fiber, panels,leads and/or a pad which emits electrons and/or photons, such as aradiofrequency waveform, an electron current, laser photons, LEDphotons, and/or broadband light photons having a waveform length in aportion of the electromagnetic spectrum between 200 nm-2500 nmwavelength. As another example, a device and/or energy source may havean opening, a handpiece, a fiber, panels, leads and/or pad, and beconfigured to generate a radiofrequency waveform and/or electron currenthaving a wattage of less than 40 W per beam or current, and/or having atotal energy in joules of less than 40 J/cm2 per beam. As anotherexample, a device may generate a beam of photons with a wavelength rangeof between 200 nm and 2500 nm at an emitted power between 0.001 W to 50W per beam or current, and/or having irradiance between 0.001 W to 10W/cm2 per beam or current. As another example, a device may generatephotons in the wavelength range from about 200 nm to about 2500 nm, atan emitted power between 0.001 W to 50 W, having irradiance between0.001 W to 10 W/cm2. As another example, a device may generate an RFwaveform and/or waveforms, having an emitted power range of between0.0001 W and 5 W, 8 W or lower, or 6 W or lower; and/or a dose between 1and 50 J/cm2. As another example, a device may include a laser, LED,and/or broadband energy source configured to produce a beam and/or beamsof photons having a wavelength from about 200 nm to about 2500 nm,and/or a beam and/or beams of photons having a wavelength in the red andIR wavelength range (630 nm-1064 nm) at a power of 0.001 to 2 W perbeam, and/or having an irradiance of 1 to 300 mW/cm2.

A device may include one or more fiber optic components (or fibers). Afiber optic component may be placed in and/or proximate to treatedtissue, for example. In some embodiments, the one or more fiber opticcomponents may comprise one or more individual fibers (e.g., conduits).Individual fibers may conduct energy having the same or differentcharacteristics. For example, individual fibers may conduct energyhaving different wavelengths, and/or different powers (or averagepowers). As another example, one fiber may conduct LED light, whileanother fiber may conduct a different type of energy.

Lasers

In some embodiments, laser energy comprises a laser a beam of lighthaving a wavelength in the green wavelength range (520-570 nm), redwavelength range (620-750 nm), or yellow wavelength range (570-590 nm),having an alternative wattage of 0.001 W to 5 W and/or 0.002 W to 4 W,and/or 0.003 W to 3 W, and/or 0.005 W to 2 W per beam. In someembodiments, the laser light utilizes the IR wavelength range (700nm-1400 nm) at a laser power of 0.001 W to 5 W per beam to treat tissue.In some embodiments, a device may include a single laser that is used,with or without multiple individual fibers, with or without a handpiece,with or without a lens. In some embodiments, multiple lasers are used ina single device, with or without multiple individual fibers, with orwithout multiple lenses, with or without a handpiece or handpieces(e.g., a housing). In some embodiments, a device may include one or morelasers comprising one or more fibers (e.g., as further described belowand shown in a figure). The one or more fibers may comprise conduitsconfigured to conduct laser light (energy) from a source to an outputport or opening in a housing of a device. One or more beams may beseparated into and/or otherwise conducted by one or more fibers, forexample. In some embodiments, one or more individual fibers may beconfigured to conduct the laser radiation, where the laser radiation hasa power of about 0.1 to 5 W per fiber. A device may be configured suchthat different fibers may conduct energy having differentcharacteristics (e.g., different wavelengths, powers, frequencies,etc.). Such fibers may be used in one or more of the other devicesdescribed herein. For example, one or more such fibers may be used inconjunction with an imaging device described herein.

In some embodiments, fibers may be assembled in a line or in a cross,multi-cross or a geometric form. Precise control of what is coming outeach fiber without system loss over time may be achieved with a closeddevice. In some embodiments, a sapphire lens may be used at the distalend of a fiber handpiece to close off the device. A given fiber may beenclosed in a tubing that does not bend at a high degree. Other lensescan be used such as quartz or even plastic. This closed device does notenter the body. Another type of fiber device may be configured to enterthe body. Such a device may have disposable fibers. The attachment of adisposable tip would be a system loss. One way to avoid that is toincrease the reflectivity in the tube (fiber or handpiece/housing) tocompensate for probable loss.

By way of a non-limiting example, a specific embodiment of a laser thatmay be used in a device may have some or all of the followingcharacteristics. The example laser may comprise a red or infrared fiberlaser with varying degrees of collimation of the beam, with a maximumoutput power of 12 mW-250 mW at 635 nm-808 nm (+/−5 nm), having a fibercore diameter of 200 um/0.37 NA, with a low operating current of lessthan 480 mA or less than 70 mM, a low operating voltage of 5V DC, it maybe a class Ma laser, it may be CDRH/ROHS compatible, it may be stat,surge, and reverse polarity protected, it may be configured with an autoconstant current, have an operating temperature of −10 C to 50 C, andable to operate at up to 90% humidity. It may have a spectral width ofabout 1 nm, with a wavelength shift of about 0.2 nm. Fibercharacteristics may include a 14 (Diameter)×50 (Length) mm shape, with afiber core diameter of 200 um, a 3 mm cladding diameter, a 0.37 NA, andan SMA connector.

LED

In some embodiments, a device may include one or more LED's. In someembodiments, an LED is configured to utilize the IR wavelength range. Insome embodiments, an LED is configured to utilize one or more wavelengthranges that are part of the visible light spectrum (e.g., the greenwavelength range, the red wavelength range, the blue wavelength range,etc.) In some embodiments, a device may include a plurality of LED's.The plurality of LED's may be arranged in a pattern, for example. Theplurality of LED's may be included in a LED board, and/or othercomponents. For example, the plurality of LED's may include enough LED'sto substantially cover a board of a given size (e.g., a 12 inch×12 inchboard, a 7 inch×12 inch board, etc.) In some embodiments, LED's may bearranged in a symmetrical array (e.g., a 2×2 array, a 3×3 array, a 4×4array, a 5×5 array, a 6×6 array, a 7×7 array, an 8×8 array, a 9×9 array,a 10×10 array, an 11×11 array, a 12×12 array, etc.), in non-symmetricalarrays (e.g., 1×2, 3×5, 4×5, etc.), and/or have other arrangements. Insome embodiments, an LED board may be configured to be held by a user(e.g., so that the energy from the LED's can be directed toward tissue),self-supporting (e.g., an LED board can form a stand that can be placednext to tissue and stand up right), and/or have other configurations.These examples are not intended to be limiting.

RF/Audio

In some embodiments, RF or audio frequency energy may have a power of upto 10 W or 80 Vpp per wave (volts peak to peak per wave). An RF carrierwave frequency may be in the range of 0.1 MHz to 20 MHz, while anon-sinusoidal waveform may be in the range of 0.5 to 40 KHz, or from0.5 to 24 GHz. In some embodiments, the carrier wave frequency is in therange of 0.2 MHz to 10 MHz, and/or 0.3 MHz to 5 MHz. In someembodiments, a 0.001 W to 10 W range RF or audio frequency energy,and/or a 0.001 W to 3 W range, is utilized in the hertz range of 40 Hzto 24 GHz. In some embodiments, RF or audio frequency comprises morethan one sinusoidal or non-sinusoidal wave wherein the more than onewave may demonstrate a pattern, such as but not limited to a harmonicspattern. Each wave in the pattern may have the power parametersdescribed herein, for example.

In some embodiments, RF or audio frequency may have a power of up to 10W per opening and be comprised of a carrier wave have a frequency in therange of 0.1 MHz to 20 MHz and a non-sinusoidal waveform in the range of0.5 to 40 KHz. In some embodiments, the carrier wave frequency is in therange of 0.2 MHz to 10 MHz, and/or 0.3 MHz to 5 MHz. In someembodiments, a 0.001-10 W range RF or audio frequency, and/or a 0.001 Wto 3 W range, is utilized in the hertz range of 40 Hz to 24 GHz. In someembodiments, the RF or audio frequency comprises more than one wavewherein at least one wave demonstrates a harmonics pattern. In someembodiments, the at least one non-sinusoidal waveform may be in therange of the above parameters as single or multiple waveforms in thepresence or absence of a carrier wave.

In some embodiments, audio energy may be generated by the samecomponents/device that generate the RF energy. Audio waves are justlower frequency waves compared to the RF waves. Audio frequency wavesmay be generated by a similar device having a handpiece, a pad, a lead,a bar, a probe, and/or other components configured to generate sound(audio waves/energy.)

In some embodiments, RF and/or audio energy may be generated and/orcontrolled at least in part using a raspberry pi computing module and/orother devices. An example of such a device may be sold by Digi-KeyElectronics(https://www.digikey.com/en/product-highlight/r/raspberry-pikaspberry-pi-4-modelb?utm_adgroup=xGeneral&utm_source=google&utm_medium=cpc&utm_campaign=Dynamic%20Search&utm_term=&utm_content=xGeneral&gclid=CjwKCAjwjqT5BRAPEiwAJ1BuBVHEruloeGpdsl0LIV6-aZvQaZezuiiL9NDJfi2N_WSa7bG63w9OoBoCysYQAvD_BwE)and/or other providers. Such a device may be a single board computerwith a 64 bit quad core processor, dual display support, HDMI ports, 8GB of RAM, dual band 2.4/5.0 GHz wire LAN components, Bluetoothcomponents, and/or other components. In some embodiments, such a devicemay be user programmed to generate energy having the characteristicsdescribed herein.

Imaging

In some embodiments, an imaging device may be included in one or more ofthe present systems and/or used in one or more of the present methods.An imaging device may be configured to obtain and/or analyze images oftissue and vascular structures at or beneath the epithelium and/or othertissue. The imaging device may or may not have a handle (e.g.,embodiments may include a stand and/or other supporting components). Insome embodiments, an image device head (which is coupled to the handle)includes a camera, one or more LED's and/or other components. In someembodiments, the one or more LED's that are included in an imagingdevice may operate at the same or similar wavelengths, powers,frequencies, etc. described herein for the various types of energy. Animaging device may include a display screen, a user entry device (e.g.,a keyboard), one or more processors configured to analyze imagesobtained by the camera and/or other information, and/or othercomponents. In some embodiments, the handle and the head may beconfigured to be moved back and forth over an area of tissue that isbeing illuminated by the LED's (e.g., producing visible light in thegreen wavelength range) and/or imaged by the camera. The camera mayacquire a plurality of images during this movement. The one or moreprocessors may analyze and combine the images to produce a threedimensional image of the tissue and/or features within the tissue. Theone or more processors may be configured to “stack” the images, forexample.

In some embodiments, an imaging device may facilitate finding veins intissues. The imaging device may produce at least one output indicatingthe location of veins for the application of tissue analysis in woundtreatment. The tissue may be illuminated by an optical probe (e.g.,including the components described above). Reflected light is capturedby the camera (e.g., having sensors sensitive to different wavelengthsof light), which is connected to a computer (e.g., including the one ormore processors). In some embodiments, the optical probe may beconfigured to illuminate the tissue with infrared and/or green lights.In some embodiments, the image stacking and/or other analysis by the oneor more processors may facilitate 3d image visualization, machinelearning or deep learning algorithm vein detection, edge detection,and/or other operations. In some embodiments, the one or more processorsmay be, may be included in, and/or may form a raspberry pi computingmodule (e.g., as described above).

The following figures illustrate several non-limiting examples of thedevices and/or device components described above.

FIG. 21 is an electrical drawing of an example of a function generatorthat may be configured to generate audio and/or radiofrequency signalsas described herein. FIG. 22 is a mechanical drawing of an example of afunction generator configured to generate audio and/or radiofrequencysignals. The function generator may be considered an energy source, forexample. As shown in FIG. 21, the function generator includes variousresistors, grounds, capacitors, and other components arranged as shown.In some embodiments, as described above, the audio and/or radiofrequencyenergy (e.g., the audio and/or radiofrequency waves/signals) may begenerated by a programmed raspberry pi computing module (e.g., formed byand/or including one or more processors). The components of the audioand/or radio frequency energy source (e.g., a function generator, araspberry pi computing module, etc.) may be selected and/or arranged toproduce an audio or RF wave with a sine wave frequency range of about 1Hz-500 kHz, a rectangle wave frequency range of about 1 Hz-20 kHz, atriangle wave frequency range of about 1 Hz-20 kHz, or a sawtooth wavefrequency range of about 1 Hz-20 kHz, for example. The components of thefunction generator may be selected and/or arranged such that thefunction generator has an output voltage of about OV to +/−10V peak topeak, with an output impedance of about 50 ohms, for example. Thecomponents of the function generator may include a microcontroller suchas an ATMega48, 8 bit microcontroller, and/or be associated with asupply voltage of about 3.5V DC-10V DC, a supply current of about 300mA, and/or other characteristics.

FIG. 23 illustrates an example LED board 100 that includes a pluralityof LEDs 102 that can be used to treat and/or image tissue, and/or beused for other purposes. In some embodiments, LED board 100 and/or LED's102 can be included in one or more devices and/or be used for one ormore methods described herein. In the example shown in FIG. 23, LED's102 are arranged in a non-symmetric 4×5 array. This is just one possibleexample of such an arrangement. In addition, board 100 may have anydimensions that allow a device comprising LED's 102 to function asdescribed herein (e.g., to therapeutically treat and/or image tissue).

FIG. 24a-24c show various views of a possible embodiment of a device 200configured to generate radiofrequency and/or audio frequency energy.FIG. 24a shows a front perspective view. FIG. 24b shows a right sideview. FIG. 24c shows a bottom view. FIG. 25a-25d show views of the RF oraudio frequency device 200 with and without a laser energy source. FIG.25a shows an exploded view of RF or audio frequency device with laser.FIG. 25b shows a close-up of the handpiece with laser relative tohousing. FIG. 25c shows an exploded view of RF or audio frequency device200 without laser. FIG. 25d shows a detailed view of a housing for an RFor audio frequency device without a laser. Device 200 may be comprisedof wire grommet 201 integrally connected to handle 209, handle 209further comprised of heat sink 202. Housing 203 securely connects toheat sink 202 thereby creating a cavity between the housing 203 and heatsink 202. Laser 204 is positioned within the cavity between housing 203and heat sink 202. Male connectors 205 connect RF source 208 to housing203 wherein threaded inserts 206 cover the connection there between.FIG. 25b shows a detailed view of laser 204 and housing 203 in relationto each other.

FIG. 26 illustrates a flow chart for an example method for using an RFor audio frequency device (e.g., device 500 and/or 200 described above).

FIGS. 27a-27e show various views of another example embodiment of an RFor audio frequency device. FIG. 27a shows a top view. FIG. 27b shows aside view. FIG. 27c shows a perspective view of the RF or audiofrequency tips. FIG. 27d shows an exploded side perspective view. FIG.27e shows an alternative side perspective view. A device 500 for use inconjunction with the systems, compositions, and methods of the presentinvention is shown. FIG. 27a shows a top view of the device 500. FIG.27b shows a side view and FIG. 27c shows a close-up of the tip of device500. Specifically, FIG. 27d illustrates an exploded view of the device500 comprised of housing 503, tips 505 and energy source 507. Energysource 507 provides RF or audio frequency in this example. FIG. 27eshows a side perspective view of the assembled device 500.

FIG. 28 is an electrical drawing of example circuitry that may beincluded in a device such as device 200 and/or 500 described above. Forexample, the circuitry may include a power source, various wires (e.g.,labeled by colors), a laser diode, electrodes (e.g., for outputting RFand/or audio energy), and/or other components.

FIG. 29 is a schematic illustration of an example device that uses laserenergy for treatment of tissue. The example device includes a light pipe1, laser emitting diodes 2 for pumping, a laser delivery means adapter 3(e.g., any sort of part or component than can be used to couple thelaser delivery means (described next), such as a flexible, adjustable,and/or otherwise moveable portion of a housing and/or a component thatcan be coupled to a housing, etc.), a laser delivery means 4 (e.g., afiber and/or other conduits), an optics instrument 5 configured forinteraction with a patient (e.g., a quartz lens, a sapphire lens, ahousing, etc.), a feedback controlled substantially constant currentsource 6, a plug in 7 to connect a laser to a driver, a cylindricalwaveguide with a mirror coating 8 (e.g., see the fiber in the nextfigure), cavity mirrors 9, and/or other components. In some embodiments,a device that uses laser energy may include more, less, or differentcomponents than those shown in FIG. 29. FIG. 29 is an illustration of asingle laser. In some embodiments, a single laser is used in a device,with or without a fiber (described below), with or without a handpiece(e.g., a housing or handle), with or without a lens. In someembodiments, multiple lasers are used in a single device, with orwithout multiple fibers, with or without multiple lenses, with orwithout a handpiece or handpieces.

FIG. 30 illustrates an example of laser fibers (e.g., conduits, fiberoptic components) 110 that may be included in a device that uses laserenergy for treatment of tissue. Possible components and/or dimensionsassociated with such fibers are listed in FIG. 30. These possiblecomponents and/or dimensions are examples only, and may be varied fromwhat is shown in FIG. 30, provided laser fibers 110 still allow the oneor more devices describe herein to function as intended. In someembodiments, a given fiber may comprise a (e.g., pure) fused silicacore, fluorine doped fused silica cladding, a jacket, and/or othercomponents. A given fiber may be configured to conduct laser energy froma laser energy source so that it can be directed at tissue fortreatment. In some embodiments, a laser fiber as shown in FIG. 30 mayform one or more of the components shown in FIG. 29.

FIG. 31 illustrates combining individual laser fibers for a device thatuses laser energy for treatment of tissue. FIG. 31 is an enlarged viewof a ferrule shown in FIG. 30. As shown in FIG. 31, multiple individuallaser fibers may be surrounded by a ferrule (having two parts in thisexample). The individual fibers extend from a window, through a bolster,and are collected so that they can pass through a protection tube. Insome embodiments, the window may be silica for example. In someembodiments, one or more fibers may be configured and/or use such that aforce and/or effect of light and/or other energy is not diminished bydistribution of the light along a line or pattern. If a longer beam wasused, the force would be diminished at the distal ends of the beam, forexample. Also the patterns seen in the tissue can be matched by usingspecific patterns of fibers with specific arrangements, orientations,etc.

FIG. 32 illustrates an example of components of a possible embodiment ofan LED device that may be included in a present system and/or used in apresent method. For example, FIG. 32 illustrates an LED waveguide 701, aplug in 702 configured to connect the LED to a driver, a feedbackcontrolled substantially constant current source 703, cavity mirrors704, multi-layer semiconductor materials 705, wire bonding means 706, awavelength filter 707 (e.g., a sapphire or silica lens), an LED todelivery means adaptor 708, LED delivery means 709, an optics instrument710 configured to facilitate interaction with a patient, and/or othercomponents. In some embodiments, cavity mirrors 704, wavelength filter707, LED to delivery means adaptor 708, LED delivery means 709, andoptics instrument 710 are optional components. In some embodiments,multiple LED's (e.g., including the components shown in FIG. 32) may beused in a single device.

FIG. 33 illustrates another example of components of a possibleembodiment of an LED device that may be included in a present systemand/or used in a present method. For example, FIG. 33 illustrates an LEDwaveguide 801, a plug in 802 configured to connect the LED to a driver,a feedback controlled substantially constant current source 803, cavitymirrors 804, multi-layer semiconductor materials 805, wire bonding means806, a wavelength filter 807 (e.g., a sapphire or silica lens), an LEDto delivery means adaptor 808, LED delivery means 809, an opticsinstrument 810 configured to facilitate interaction with a patient,and/or other components. In some embodiments, cavity mirrors 804,wavelength filter 807, LED to delivery means adaptor 808, LED deliverymeans 809, and optics instrument 810 are optional components. In someembodiments, multiple LED's (e.g., including the components shown inFIG. 33) may be used in a single device.

FIG. 34 illustrates an example of components of an imaging device thatmay be included in a present system and/or used in a present method.FIG. 34 illustrates an imaging device having a handle 902 (e.g., someembodiments may include a stand and/or other supporting components). Insome embodiments, an image device head 904 (which is coupled to handle902) includes a camera, one or more LED's and/or other components. Animaging device may include a display screen 906, a user entry device 908(e.g., a keyboard), one or more processors configured to analyze 910images 912 obtained by the camera and/or other information, and/or othercomponents. In some embodiments, the handle and the head may beconfigured to be moved back and forth over an area of tissue that isbeing illuminated by the LED's and/or imaged by the camera. The cameramay acquire a plurality of images during this movement. The one or moreprocessors may analyze and combine the images to produce a threedimensional image 914 of the tissue and/or features within the tissue.In this example, the images and/or the analysis may be used to detectcirculation blockages, circulation loss, tissue density changes, and/orother conditions.

In some embodiments, the one or more processors may be programmed withPython, C++, and/or other programming code. The one or more processorsmay be configured to determine how far tissue is from a focal pointassociated with a vein, and provide visualization of the distance on adisplay. The one or more processors may be configured to cause thecamera to take multiple images when the tissue distance becomes close tothe focal point, and cause the display to indicate when images aretaken. The one or more processors may be configured to store dataelectronically according to a specific protocol, communicate data (e.g.,to a doctor) also with a specific protocol, and/or cause display of thedata on a remote computing device (e.g., a doctor's computer) such thatthe data may be visualized. In some embodiments, the one or moreprocessors are configured to execute machine learning algorithms toextract data from the images.

The terms “power” and “average power” may be used interchangeablyherein, with “power” referring to continuous power applied to lasers,LEDs broadband light, radiofrequency, audio or current devices, while“average power” refers to pulsed power applied to lasers, LEDs,broadband light, radiofrequency or current devices of a device or powersource.

Similarly, as used herein, the terms “photons” and “photonic electrons”are considered synonymous. The terms “electrons” and “photonicelectrons” are considered synonymous. The terms “lines” and “pathways”are considered synonymous. The terms “protocol” and “method” areconsidered synonymous. The terms “waveform length” and “wavelength” areconsidered synonymous. The terms, “electron current” and “electriccurrent” are considered synonymous.

Compositions

In some embodiments, provided herein are compositions for use inconducting a method disclosed herein. Compositions are sometimesreferred to herein as substrates. A composition as described herein maybe a liquid, gel, paste, or powder, for example. In some embodiments, acomposition is substantially dry (e.g., in a fibrous or powdered form).In certain embodiments, a dry composition comprises a water content ofless than 5%, less than 2%, less than 1%, less than 0.5% or less than0.1%. In certain embodiments, a dry composition comprises an alcohol ororganic solvent content of less than 5%, less than 2%, less than 1%,less than 0.5% or less than 0.1%.

Collagen

In some embodiments, a composition comprises collagen. In certainembodiments, collagen comprises collagen fibers. In some embodiments,collagen is partially or completely hydrolyzed collagen. In someembodiments, collagen is an acid treated collagen. In some embodiments,collagen comprises a limed collagen or collagen subjected to a limingprocess. In some embodiments, collagen is provided as a collagencomposition comprising collagen and tricalcium phosphate. In certainembodiments, a collagen composition comprises collagen and poroustricalcium phosphate crystals. Porous tricalcium phosphate crystals maycomprise a single size of crystals or different sizes of crystals nonlimiting examples of which include tricalcium phosphate crystals havinga mean, average or absolute diameter in a range of 4 μm to 6 mm, 4-50μm, 50-150 μm, 100-300 μm, 500-1000 μm, 1-3 mm, 3-6 mm, or intermediateranges thereof.

In some embodiments, a composition comprises collagen in an amount in arange of 0.1-70% wt/wt (wt of collagen/total wt of composition). In someembodiments, a composition comprises collagen in an amount in a range of0.1-70% (wt/dry wt) (wt of collagen/total dry wt of composition). Insome embodiments, a composition comprises collagen in an amount in arange of 30-70%, 40-65% or 50-60% (wt/wt). In some embodiments, acomposition comprises collagen in an amount of about 50-60% (wt/wt) or50-60% (wt/dry wt). In some embodiments, a composition comprises purecollagen.

In some embodiments, a composition comprises collagen in an amount in arange of 0.01-10%, 0.01-5%, 0.01-1%, 0.05-0.5% or 0.05-0.12% (wt/wt). Insome embodiments, a composition comprises collagen in an amount of about0.1% (wt/wt). In some embodiments, a composition disclosed herein doesnot comprise collagen.

Hyaluronic Acid

In some embodiments, a composition comprises a suitable hyaluronic acid(HA). In some embodiments, a composition comprises HA in an amount in arange of 10% to 80%, 20% to 40%, 25% to 35%, or 26% to 30% (wt/wt). Insome embodiments, a composition comprises HA in an amount in a range of10% to 40%, 20% to 40%, 25% to 35%, or 26% to 30% (wt/dry wt). In someembodiments, a composition comprises HA in an amount of 27-30% (wt/drywt).

In some embodiments, a composition comprises HA in an amount in a rangeof 0.01%-10%, 0.1% to 5%, 0.1% to 1%, 0.2% to 0.9%, or 0.3% to 0.8%(wt/wt) or (wt/vol). In some embodiments, a composition comprises HA inan amount in a range of 0.5%-0.6 (wt/wt).

In some embodiments, a composition disclosed herein does not comprisehyaluronic acid.

Sugars

In some embodiments, a composition comprises one or more sugars,non-limiting examples of which include mannose, galactose,N-acetylglactosamine, N-acetylglucosamine, N-acetylneuraminic acid,fucose, xylose and combination thereof. In some embodiments, acomposition comprises a sugar in an amount in a range of 0.001%-35%. Insome embodiments, a composition comprises a sugar in an amount in arange of about 0.001% to 5%, about 0.005% to 5%, about 0.01% to 0.5% orabout 0.05-0.2% (wt/vol). In some embodiments, a composition comprises asugar in an amount in a range of about 5%-35%, 10%-30%, 15%-30% or20-25% (wt/wt) or (wt/dry wt).

In some embodiments, a composition comprises fucose. In someembodiments, a composition comprises fucose in an amount in a range of0.01%-25%, 0.05%-15%, 0.05% to 2% or 8-15% (wt/wt), or intermediateranges thereof.

Metals

In some embodiments, a composition comprises a metal, or a salt thereof,non-limiting examples of which include copper (Cu), gold (Au), silver(Ag), platinum (Pt), iron (Fe), zinc, selenium, manganese, cobalt,chromium, boron, molybdenum, silicon, nickel, vanadium and combinationsthereof. In some embodiments, a composition comprises one or moremetals, or a suitable salt thereof, in an amount in a range of0.00001-10%, 0.0001-10%, 0.0001-5%, 0.001-5%, 0.01-5%, 0.5%-15%, 1%-10%,1%-5%, 2%-3% (wt/wt) or intermediate ranges thereof. In someembodiments, a composition comprises one or more metals, or saltsthereof, each at a concentration in a range of about 1% to 15%, about 2%to 10%, or about 5% to 10% (wt/dry wt). In some embodiments, one or moreof these may be in ionic form.

In some embodiments, a composition comprises one or more metals, each ata concentration in a range of about 0.001% to 1%, about 0.01% to 1%, orabout 0.01% to 0.1% (wt/wt). In some embodiments, a composition is a wetcomposition (e.g., a paste, gel or liquid) and comprises one or moremetals, each at a concentration in a range of about 0.001% to 1%, about0.01% to 1%, or about 0.01% to 0.1% (wt/vol). In some embodiments, ametal, or salt thereof, is present in a composition in the form of aparticle or nanoparticle. In some embodiments, a composition comprisescopper (Cu), gold (Au), silver (Ag), platinum (Pt) and/or, iron (Fe).

In some embodiments, a suitable salt of Cu is selected from CuCl, CuCl₂and CuCl₃. In some embodiments, a composition comprises Cu, or asuitable salt thereof, in an amount in a range of 0.001-15%, 0.001%-10%,0.001%-5%, 0.001%-0.1%, 0.01-1%, 0.5%-15%, 1%-10%, 1%-5%, 2%-3%, orintermediate ranges thereof. In some embodiments, a compositioncomprises Cu, or a suitable salt thereof, in an amount in a range of0.01-1%.

In some embodiments, a suitable salt of Au is selected from AuCl, AuCl₂,AuCl₃ and Au₂O₃. In some embodiments, a composition comprises Au, or asuitable salt thereof, in an amount in a range of 0.001-15%, 0.001%-10%,0.001%-5%, 0.001%-0.1%, 0.01-1%, 0.5%-15%, 1%-10%, 1%-5%, 2%-3%, 4%-10%(wt/wt) or (wt/dry wt), or intermediate ranges thereof.

In some embodiments, a suitable salt of silver (Ag) is selected fromAgCl, AgCl₂ and AgNO₃. In some embodiments, a composition comprises Ag,or a suitable salt thereof, in an amount in a range of 0.001-15%,0.001%-10%, 0.001%-5%, 0.001%-0.1%, 0.01-1%, 0.5%-15%, 1%-10%, 1%-5%,2%-3%, 4%-10% (wt/wt) or (wt/dry wt), or intermediate ranges thereof.

In some embodiments, a suitable salt of platinum (Pt) is selected fromPtCl₂, PtCl₄, PtBr₂ and Ptl₂. In some embodiments, a compositioncomprises Pt, or a suitable salt thereof, in an amount in a range of0.001-15%, 0.001%-10%, 0.001%-5%, 0.001%-0.1%, 0.01-1%, 0.5%-15%,1%-10%, 1%-5%, 2%-3%, 4%-10% (wt/wt) or (wt/dry wt), or intermediateranges thereof.

In some embodiments, a suitable salt of iron (Fe) is selected from FeCl₃or FeCl₂. In some embodiments, a composition comprises Fe, or a suitablesalt thereof, in an amount in a range of 0.001-15%, 0.001%-10%,0.001%-5%, 0.001%-0.1%, 0.01-1%, 0.5%-15%, 1%-10%, 1%-5%, 2%-3%, 4%-10%(wt/wt) or (wt/dry wt), or intermediate ranges thereof.

Salts & Electrolytes

In some embodiments, a composition comprises one or more salts orelectrolytes, non-limiting examples of which include calcium chloride,choline chloride, magnesium sulfate, potassium chloride, potassiumphosphate (monobasic), sodium bicarbonate, sodium chloride, sodiumiodide, the like or combinations thereof.

Nucleotides

In some embodiments, a composition comprises one or more nucleotides,non-limiting examples of which include ADP, ATP, cAMP, CTP, TTP, GTP,pppGpp, the like and combinations thereof. In some embodiments, acomposition comprises ATP. In some embodiments, a composition comprisesa nucleotide in an amount in a range of 0.001-35% (wt/wt or wt/dry wt).

In some embodiments, a composition comprises a nucleotide in an amountin a range of 0.001%-10%, 0.001%-1%, 0.01%-1%, 0.05-0.3% or 0.05-0.15%(wt/wt) or intermediate ranges thereof. In some embodiments, acomposition is a wet composition (e.g., a paste, gel or liquid) andcomprises a nucleotide in an amount in a range of 0.001%-10%, 0.001%-1%,0.01%-1%, 0.05-0.3% or 0.05-0.15% (wt/vol) or intermediate rangesthereof. In some embodiments, a composition comprises a nucleotide in anamount in a range of 5%-35%, 10%-30%, 15%-30% or 20-25% (wt/dry wt). Insome embodiments, a composition is a dry composition (e.g., powder, afibrous mass) and comprises a nucleotide in an amount in a range of5%-35%, 10%-30%, 15%-30% or 20-25% (wt/dry wt).

In some embodiments, a composition comprises ATP in an amount in a rangeof 0.001%-10%, 0.001%-1%, 0.01%-1%, 0.05-0.3% or 0.05-0.15% (wt/wt) orin an amount in a range of 5%-35%, 10%-30%, 15%-30% or 20-25% (wt/drywt).

In some embodiments, a composition comprises one or more free bases,non-limiting examples of which include adenosine, uridine, guanosine,iridine and cytidine. In some embodiments, a composition comprises oneor more free bases each in an amount in a range of 0.001-35% (wt/wt orwt/dry wt).

Acetylcholine

In some embodiments, a composition comprises acetylcholine (ACh). Insome embodiments, a composition comprises ACh in an amount in a range of0.001%-10%, 0.001%-1%, 0.01%-1%, 0.05-0.3% or 0.05-0.15% (wt/wt) orintermediate ranges thereof. In some embodiments, a composition is a wetcomposition (e.g., a paste, gel or liquid) and comprises ACh in anamount in a range of 0.001%-10%, 0.001%-1%, 0.01%-1%, 0.05-0.3% or0.05-0.15% (wt/vol) or intermediate ranges thereof. In some embodiments,a composition comprises ACh in an amount in a range of 5%-35%, 10%-30%,15%-30% or 20-25% (wt/dry wt). In some embodiments, a composition is adry composition (e.g., powder, a fibrous mass) and comprises ACh in anamount in a range of 5%-35%, 10%-30%, 15%-30% or 20-25% (wt/dry wt).

Acids

In some embodiments, a composition comprises an acid. In someembodiments, a composition comprises hydrochloric acid (HCl) orglycine-HCL in an amount in a range of 0.0001-60%, 0.0001%-10%,0.0001%-5%, 0.0001%-1%, 0.0001%-0.1%, 0.0005%-0.1%, 0.001-0.1%, 5%-15%,1%-10%, 1%-5%, 2%-3% (wt/wt) or intermediate ranges thereof. In someembodiments, a composition comprises hydrochloric acid (HCl) orglycine-HCL in an amount in a range of 0.0001-1.0%, 0.0001-0.75%,0.0005-0.75%, or 0.0005-0.5% (wt/wt).

In some embodiments, a composition comprises a pH in a range of 2 to 8,3 to 7, 3.5-7 or 4 to 6. In some embodiments, a composition comprises apH in a range of 2-7, 2-6, 2-5, 2-4, 2-3 or 2.5-4. In some embodiments,a composition is a dry composition (e.g., a powder or fibrous mass) andcomprises a pH in a range of 1-4, 1.5-3.5 or 2-3. In some embodiments, acomposition is a wet composition (e.g., a slurry, gel, paste or liquid)and comprises a pH in a range of 1-8, 2-8 or 3-7.

In some embodiments, a composition comprises benzoic acid. In someembodiments, a composition comprises benzoic acid in an amount in arange of 0.001-25%, 0.001%-10%, 0.001%-5%, 0.001%-0.1%, 0.01-1%,0.5%-15%, 1%-10%, 1%-5%, 2%-3% (wt/wt) or intermediate ranges thereof.

Water

In some embodiments, a composition comprises a water content in a rangeof 0%-90%, 0% to 20%, or 0% to 10%, or 0% to 5% (wt/wt). In someembodiments, a composition is substantially dry (e.g., a powder,granules, a fibrous mass) and comprises a water content in a range ofless than 5%, less than 2%, less than 1%, less than 0.5% or less than0.1%. In certain embodiments, a composition is substantially dry (e.g.,a powder, granules, a fibrous mass) and comprises an alcohol or organicsolvent content of less than 5%, less than 2%, less than 1%, less than0.5% or less than 0.1%.

Hydroxyapatite

In some embodiments, a composition comprises hydroxyapatite.Hydroxyapatite may be of a suitable size and a suitable form. In someembodiments, a composition comprises hydroxyapatite in the form ofcrystals which may be dense or porous.

Amino Acids

In some embodiments, a composition comprises one or more amino acids. Insome embodiments, a composition comprises one or more essential aminoacids. In some embodiments, a composition comprises lysine, proline,isoleucine, leucine, methionine, phenylalanine, threonine, tryptophan,valine and/or histidine. In some embodiments, a composition comprisesone or more non-essential amino acids, non-limiting examples of whichinclude alanine, arginine, aspartate, glutamate, glycine, serine andproline.

Fatty Acids

In some embodiments, a composition comprises one or more fatty acids,non-limiting examples of which include linoleic acid (LA) and/oralpha-linolenic acid (ALA).

Vitamins

In some embodiments, a composition comprises one or more vitamins,non-limiting examples of which include vitamin A, vitamin D2, vitaminD3, vitamin E, vitamin Ki, vitamin K2, vitamin B1, vitamin B2, vitaminB3, vitamin B5, vitamin B6, vitamin B7, vitamin B9, vitamin C andvitamin B12 (methylcobalamin, hydroxocobalamin).

Other Supplements

In some embodiments, a composition comprises cholesterol, diacylglycerol(DAG), diglycerol, and/or pantothenic acid.

Exemplary Compositions

In some embodiments, a composition comprises one or more ingredientsselected from collagen, hyaluronic acid, a metal or salt thereof, asugar (e.g., fucose), a nucleotide (e.g., ATP), ACh, water, an acid,fucose, an amino acid, a vitamin, a fatty acid, hydroxyapatite, and anelectrolyte. In some embodiments, a composition comprises HA, and one ormore of collagen, fucose, copper, and iron. In some embodiments, acomposition comprises HA, and one or more of collagen, fucose, copper,and iron.

In some embodiments, a composition comprises a formulation shown in anyone of FIGS. 35-47. In some embodiments, a composition compriseshyaluronic acid (HA)(Formula 1). In some embodiments, a compositioncomprises hyaluronic acid (HA) and fucose (Formula 2). In someembodiments, a composition comprises hyaluronic acid (HA) and copper(Formula 3). In some embodiments, a composition comprises hyaluronicacid (HA) and iron (Formula 4). In some embodiments, a compositioncomprises hyaluronic acid (HA) and silver (Formula 5). In someembodiments, a composition comprises hyaluronic acid (HA) and gold(Formula 6). In some embodiments, a composition comprises HA andcollagen (Formula 7). In some embodiments, a composition comprises HAand ATP (Formula 8). In some embodiments, a composition comprises HA andACh (Formula 9). In some embodiments, a composition comprises HA, fucoseand ATP (Formula 10). In some embodiments, a composition comprises HA,fucose and ACh (Formula 11). In some embodiments, a compositioncomprises HA, fucose and copper (Formula 12). In some embodiments, acomposition comprises HA, fucose and iron (Formula 13). In someembodiments, a composition comprises HA, fucose and silver (Formula 14).In some embodiments, a composition comprises HA, fucose and gold(Formula 15). In some embodiments, a composition comprises HA, copperand gold (Formula 16). In some embodiments, a composition comprises HA,collagen, copper and gold (Formula 17). In some embodiments, acomposition comprises HA, collagen, copper, gold and ATP (Formula 18).In some embodiments, a composition comprises HA, collagen, copper, gold,ATP and iron (Formula 19). In some embodiments, a composition comprisesHA, collagen, copper, gold, ATP, iron and silver (Formula 20). In someembodiments, a composition comprises HA, fucose, collagen, copper, gold,ATP, iron and silver (Formula 21). In some embodiments, a compositioncomprises fucose (Formula 22). In some embodiments, a compositioncomprises collagen (Formula 23).

In some embodiments, a composition comprises HA in an amount of 0.3-1.2%(wt/vol); optionally collagen in an amount of 0.05-0.2% (wt/vol); ironor a salt thereof in an amount of 0.015-0.06% (wt/vol) and copper or asalt thereof in an amount of 0.015-0.06% (wt/vol). In some embodiments,a composition is a wet composition formulated for topical or localadministration and comprises HA in an amount of 0.3-1.2% (wt/vol);optionally collagen in an amount of 0.05-0.2% (wt/vol); iron or a saltthereof in an amount of 0.015-0.06% (wt/vol) and copper or a saltthereof in an amount of 0.015-0.06% (wt/vol).

In some embodiments, a composition comprises collagen in an amount of50-70% (wt/wt); optionally HA in an amount of 25-35% (wt/wt); iron or asalt thereof in an amount of 4.5-10% (wt/wt) and copper or a saltthereof in an amount of 4.5-10% (wt/wt). In some embodiments, acomposition is a substantially dry composition formulated for topical orlocal administration and comprises collagen in an amount of 50-70%(wt/wt); optionally HA in an amount of 25-35% (wt/wt); iron or a saltthereof in an amount of 4.5-10% (wt/wt) and copper or a salt thereof inan amount of 4.5-10% (wt/wt).

In some embodiments, a composition does not comprise collagen. In someembodiments, a composition does not comprise HA. In some embodiments, acomposition does not comprise fucose. In some embodiments, a compositiondoes not comprise copper. In some embodiments, a composition does notcomprise silver. In some embodiments, a composition does not compriseiron. In some embodiments, a composition does not comprise gold. In someembodiments, a composition does not comprise ATP. In some embodiments, acomposition does not comprise ACh.

Pharmaceutical Compositions & Formulations

In some embodiments, a pharmaceutical composition comprises atherapeutically effective amount of a composition disclosed herein. Insome embodiments provided herein is a pharmaceutical composition use inconducting a method described herein. In some embodiments, apharmaceutical composition comprises a pharmaceutically acceptableexcipient, diluent, additive or carrier.

A pharmaceutical composition can be formulated for a suitable route ofadministration. In some embodiments a pharmaceutical composition isformulated for oral, subcutaneous (s.c.), intradermal, intramuscular,intraperitoneal and/or intravenous (i.v.) administration. In certainembodiments, a pharmaceutical composition contains formulation materialsfor modifying, maintaining, or preserving, for example, the pH,osmolarity, viscosity, clarity, color, isotonicity, odor, sterility,stability, rate of dissolution or release, adsorption or penetration ofthe composition. In certain embodiments, suitable formulation materialsinclude, but are not limited to, amino acids (such as glycine,glutamine, asparagine, arginine or lysine); antimicrobials; antioxidants(such as ascorbic acid, sodium sulfite or sodium hydrogen-sulfite);buffers (such as borate, bicarbonate, Tris-HCl, citrates, phosphates(e.g., phosphate buffered saline) or suitable organic acids); bulkingagents (such as mannitol or glycine); chelating agents (such asethylenediamine tetraacetic acid (EDTA)); complexing agents (such ascaffeine, polyvinylpyrrolidone, beta-cyclodextrin orhydroxypropyl-beta-cyclodextrin); proteins (such as serum albumin,gelatin or immunoglobulins); coloring, flavoring and diluting agents;emulsifying agents; hydrophilic polymers (such as polyvinylpyrrolidone);low molecular weight polypeptides; salt-forming counter ions (such assodium); solvents (such as glycerin, propylene glycol or polyethyleneglycol); diluents; excipients and/or pharmaceutical adjuvants. Inparticular, a pharmaceutical composition can comprise any suitablecarrier, formulation, or ingredient, the like or combinations thereof aslisted in “Remington: The Science And Practice Of Pharmacy” MackPublishing Co., Easton, Pa., 19^(th) Edition, (1995)(hereafter,Remington '95), or “Remington: The Science And Practice Of Pharmacy”,Pharmaceutical Press, Easton, Pa., 22^(nd) Edition, (2013)(hereafter,Remington 2013), the contents of which are incorporated herein byreference in their entirety.

In certain embodiments, a pharmaceutical composition comprises asuitable excipient, non-limiting examples of which includeanti-adherents (e.g., magnesium stearate), a binder, fillers,monosaccharides, disaccharides, other carbohydrates (e.g., glucose,mannose or dextrin), sugar alcohols (e.g., mannitol or sorbitol),coatings (e.g., cellulose, hydroxypropyl methylcellulose (HPMC),microcrystalline cellulose, synthetic polymers, shellac, gelatin, cornprotein zein, enterics or other polysaccharides), starch (e.g., potato,maize or wheat starch), silica, colors, disintegrants, flavors,lubricants, preservatives, sorbents, sweeteners, vehicles, suspendingagents, surfactants and/or wetting agents (such as pluronics, PEG,sorbitan esters, polysorbates such as polysorbate 20, polysorbate 80,triton, tromethamine, lecithin, cholesterol, tyloxapol), stabilityenhancing agents (such as sucrose or sorbitol), and tonicity enhancingagents (such as alkali metal halides, sodium or potassium chloride,mannitol, sorbitol), and/or any excipient disclosed in Remington '95 orRemington 2013. The term “binder” as used herein refers to a compound oringredient that helps keeps a pharmaceutical mixture combined. Suitablebinders for making pharmaceutical formulations and are often used in thepreparation of pharmaceutical tablets, capsules and granules are knownto those skilled in the art.

In some embodiments a pharmaceutical composition comprises a suitablepharmaceutically acceptable additive and/or carrier. Non-limitingexamples of suitable additives include a suitable pH adjuster, asoothing agent, a buffer, a sulfur-containing reducing agent, anantioxidant and the like. Non-limiting examples of a sulfur-containingreducing agent include those having a sulfhydryl group (e.g., a thiol)such as N-acetylcysteine, N-acetyl homocysteine, thioctic acid,thiodiglycol, thioethanolamine, thioglycerol, thiosorbitol, thioglycolicacid and a salt thereof, sodium thiosulfate, glutathione, and a C1-C7thioalkanoic acid. Non-limiting examples of an antioxidant includeerythorbic acid, dibutyl hydroxytoluene, butylhydroxyanisole,alpha-tocopherol, tocopherol acetate, L-ascorbic acid and a saltthereof, L-ascorbyl palmitate, L-ascorbyl stearate, sodium bisulfite,sodium sulfite, triamyl gallate and propyl gallate, as well as chelatingagents such as disodium ethylenediaminetetraacetate (EDTA), sodiumpyrophosphate and sodium metaphosphate. Furthermore, diluents, additivesand excipients may comprise other commonly used ingredients, forexample, inorganic salts such as sodium chloride, potassium chloride,calcium chloride, sodium phosphate, potassium phosphate and sodiumbicarbonate, as well as organic salts such as sodium citrate, potassiumcitrate and sodium acetate.

The pharmaceutical compositions used herein can be stable over anextended period of time, for example on the order of months or years. Insome embodiments a pharmaceutical composition comprises one or moresuitable preservatives. Non-limiting examples of preservatives includebenzalkonium chloride, benzoic acid, salicylic acid, thimerosal,phenethyl alcohol, methylparaben, propylparaben, chlorhexidine, sorbicacid, hydrogen peroxide, the like and/or combinations thereof. Apreservative can comprise a quaternary ammonium compound, such asbenzalkonium chloride, benzoxonium chloride, benzethonium chloride,cetrimide, sepazonium chloride, cetylpyridinium chloride, or domiphenbromide (BRADOSOL®). A preservative can comprise an alkyl-mercury saltof thiosalicylic acid, such as thimerosal, phenylmercuric nitrate,phenylmercuric acetate or phenylmercuric borate. A preservative cancomprise a paraben, such as methylparaben or propylparaben. Apreservative can comprise an alcohol, such as chlorobutanol, benzylalcohol or phenyl ethyl alcohol. A preservative can comprise a biguanidederivative, such as chlorohexidine or polyhexamethylene biguanide. Apreservative can comprise sodium perborate, imidazolidinyl urea, and/orsorbic acid. A preservative can comprise stabilized oxychloro complexes,such as known and commercially available under the trade name PURITE®. Apreservative can comprise polyglycol-polyamine condensation resins, suchas known and commercially available under the trade name POLYQUART® fromHenkel KGaA. A preservative can comprise stabilized hydrogen peroxide. Apreservative can be benzalkonium chloride. In some embodiments apharmaceutical composition is free of preservatives.

In some embodiments a pharmaceutical composition is substantially freeof contaminants (e.g., blood cells, platelets, polypeptides, minerals,blood-borne compounds or chemicals, virus, bacteria, other pathogens,toxin, and the like). In some embodiments a pharmaceutical compositionis substantially free of serum and serum contaminants (e.g., serumproteins, serum lipids, serum carbohydrates, serum antigens and thelike). In some embodiments a pharmaceutical composition is substantiallyfree of a pathogen (e.g., a virus, parasite or bacteria). In someembodiments a pharmaceutical composition is substantially free ofendotoxin. In some embodiments a pharmaceutical composition is sterile.

The pharmaceutical compositions described herein may be configured foradministration to a subject in any suitable form and/or amount accordingto the therapy in which they are employed. For example, a pharmaceuticalcomposition configured for parenteral administration (e.g., by injectionor infusion), may take the form of a suspension, solution or emulsion inan oily or aqueous vehicle and it may contain formulation agents,excipients, additives and/or diluents such as aqueous or non-aqueoussolvents, co-solvents, suspending solutions, preservatives, stabilizingagents and or dispersing agents. In some embodiments a pharmaceuticalcomposition suitable for parenteral administration may contain one ormore excipients. In some embodiments a pharmaceutical composition islyophilized to a dry powder form. In some embodiments a pharmaceuticalcomposition is lyophilized to a dry powder form, which is suitable forreconstitution with a suitable pharmaceutical solvent (e.g., water,saline, an isotonic buffer solution (e.g., PBS), DMSO, combinationsthereof and the like). In certain embodiments, reconstituted forms of alyophilized pharmaceutical composition are suitable for parenteraladministration (e.g., intravenous administration) to a mammal.

In certain embodiments, a pharmaceutical composition is configured fororal administration and may be formulated as a tablet, microtablet,minitablets, micropellets, powder, granules, capsules (e.g., capsulesfilled with microtablets, micropellets, powders or granules), emulsions,solutions, the like or combinations thereof. Pharmaceutical compositionsconfigured for oral administration may comprise suitable coatings todelay or sustain release of the active ingredient, non-limiting examplesof which include enteric coatings such as fatty acids, waxes, shellac,plastics, methyl acrylate-methacrylic acid copolymers, cellulose acetatephthalate (CAP), cellulose acetate succinate, hydroxypropyl methylcellulose phthalate, hydroxypropyl methyl cellulose acetate succinate(hypromellose acetate succinate), polyvinyl acetate phthalate (PVAP),methyl methacrylate-methacrylic acid copolymers, cellulose acetatetrimellitate, sodium alginate, zein, plant fibers, the like andcombinations thereof.

In some embodiments a pharmaceutical compositions described herein maybe configured for topical administration and may include one or more ofa binding and/or lubricating agent, polymeric glycols, gelatins,cocoa-butter or other suitable waxes or fats. In some embodiments apharmaceutical composition described herein is incorporated into atopical formulation containing a topical carrier that is generallysuited to topical drug administration and comprising any suitablematerial known to those skilled in the art. In certain embodiments, atopical formulation of a pharmaceutical composition is formulated foradministration using a topical patch. A pharmaceutical composition, insome embodiments, is a foam, paste or gel.

In certain embodiments, an optimal pharmaceutical composition isdetermined by one skilled in the art depending upon, for example, on theintended route of administration, delivery format and desired dosage(see e.g., Remington '95 or Remington 2013, supra). A pharmaceuticalcomposition can be manufactured by any suitable manner, including, e.g.,by means of conventional mixing, dissolving, granulating, dragee-making,levigating, emulsifying, encapsulating, entrapping or tabletingprocesses (e.g., see methods described in Remington '95 or Remington2013).

Route of Administration

Any suitable method of administering a composition or pharmaceuticalcomposition disclosed herein to a subject can be used. Any suitableformulation and/or route of administration can be used foradministration of a composition disclosed herein (e.g., see Fingl et al.1975, in “The Pharmacological Basis of Therapeutics”, which isincorporated herein by reference in its entirety). A suitableformulation and/or route of administration can be chosen by a medicalprofessional (e.g., a physician) in view of, for example, a subject'srisk, age, and/or condition. Non-limiting examples of routes ofadministration include topical or local (e.g., transdermal or cutaneous,(e.g., on the skin or epidermis), in or on the eye, intranasally,transmucosal, in the ear, inside the ear (e.g., behind the ear drum)),enteral (e.g., delivered through the gastrointestinal tract, e.g.,orally (e.g., as a tablet, capsule, granule, liquid, emulsification,lozenge, or combination thereof), sublingual, by gastric feeding tube,rectally, and the like), by parenteral administration (e.g.,parenterally, e.g., intravenously, intra-arterially, intramuscularly,intraperitoneally, intradermally, subcutaneously, intracavity,intracranial, intra-articular, into a joint space, intracardiac (intothe heart), intracavernous injection, intralesional (into a skinlesion), intraosseous infusion (into the bone marrow), intrathecal (intothe spinal canal), intrauterine, intravaginal, intravesical infusion,intravitreal), the like or combinations thereof.

In some embodiments a pharmaceutical composition described herein isadministered to the lungs, bronchial passages, trachea, esophagus,sinuses, or nasal passages using a suitable method, non-limitingexamples of which include intranasal administration, intratrachealinstillation, and oral inhalative administration (e.g., by use of aninhaler, e.g., single/-multiple dose dry powder inhalers, nebulizers,and the like).

Compositions for use according to the methods of the invention can be,in some embodiments, aerosolized compositions. The aerosolizedcomposition can be formulated such that the composition has increasedsolubility and/or diffusivity. Solutions to be aerosolized can beprepared in any suitable form, for example, either as liquid solutionsor suspensions, as emulsions, or in solid forms suitable for dissolutionor suspension in liquid prior to aerosol production and inhalation. Foradministration by inhalation, the compositions described herein canconveniently be delivered in the form of an aerosol (e.g., throughliquid nebulization, dry powder dispersion or meter-dose administration.The aerosol can be delivered from pressurized packs or a nebulizer, withthe use of a suitable propellant, e.g., dichlorodifluoromethane,trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide orother suitable gas. In the case of a pressurized aerosol the dosage unitcan be determined by providing a valve to deliver a metered amount.Capsules and cartridges of, e.g., gelatin for use in an inhaler orinsufflator can be formulated containing a powder mix of a compositiondescribed herein and a suitable powder base such as lactose or starch.

For aqueous and other non-pressurized liquid systems, a variety ofnebulizers (including small volume nebulizers) can be used to aerosolizethe formulations. Compressor-driven nebulizers can utilize jettechnology and can use compressed air to generate the liquid aerosol.Such devices are commercially available from, for example, HealthdyneTechnologies, Inc.; Invacare, Inc.; Mountain Medical Equipment, Inc.;Pari Respiratory, Inc.; Mada Medical, Inc.; Puritan-Bennet; Schuco,Inc., DeVilbiss Health Care, Inc.; and Hospitak, Inc. Ultrasonicnebulizers generally rely on mechanical energy in the form of vibrationof a piezoelectric crystal to generate respirable liquid droplets andare commercially available from, for example, Omron Healthcare, Inc. andDeVilbiss Health Care, Inc. Vibrating mesh nebulizers rely upon eitherpiezoelectric or mechanical pulses to generate respirable liquiddroplets. Commercial examples of nebulizers that could be used incertain embodiments include RESPIRGARD II®, AERONEB®, AERONEB® PRO, andAERONEB® GO produced by Aerogen; AERX® and AERX ESSENCE™ produced byAradigm; PORTA-NEB®, FREEWAY FREEDOM™, Sidestream, Ventstream and I-nebproduced by Respironics, Inc.; and PARI LC-PLUS®, PARI LC-STAR®, ande-Flow7m produced by PARI, GmbH. By further non-limiting example, U.S.Pat. No. 6,196,219 is hereby incorporated by reference in its entirety.

In some embodiments a composition or a pharmaceutical compositiondisclosed herein is provided to a subject. For example, a compositionthat is provided to a subject is sometimes provided to a subject forself-administration or for administration to a subject by another (e.g.,a non-medical professional). As another example, a composition can beprovided as an instruction written by a medical practitioner thatauthorizes a patient to be provided a composition or treatment describedherein (e.g., a prescription). In yet another example, a composition canbe provided to a subject where the subject self-administers acomposition orally, intravenously or by way of an inhaler, for example.

Alternately, one can administer a composition disclosed herein in alocal rather than a systemic manner, for example, via direct applicationto a wound, skin, mucous membrane or region of interest for treating,including using a depot or sustained release formulation.

In certain embodiments a composition or pharmaceutical compositiondisclosed herein is administered alone, or is administered incombination with one or more additional active ingredients (AI) oractive pharmaceutical ingredient (API).

Dose and Therapeutically Effective Amount

In some embodiments, an amount of a composition disclosed herein (e.g.,in a pharmaceutical composition) is a therapeutically effective amount.In certain embodiments, a pharmaceutical composition comprises atherapeutically effective amount of a composition disclosed herein. Insome embodiments, a therapeutically effective amount of a compositiondisclosed herein is administered to a subject. In some embodiments, atherapeutically effective amount of a composition disclosed herein is anamount needed to obtain an effective therapeutic outcome. In certainembodiments, a therapeutically effective amount of a compositiondisclosed herein is an amount sufficient to treat a wound. Determinationof a therapeutically effective amount is well within the capability ofthose skilled in the art, especially in light of the detailed disclosureprovided herein.

In certain embodiments, a therapeutically effective amount is an amounthigh enough to provide an effective therapeutic effect (e.g., abeneficial therapeutic effect) and an amount low enough to minimizeunwanted adverse reactions. Accordingly, in certain embodiments, atherapeutically effective amount of a composition disclosed herein mayvary from subject to subject, often depending on age, weight, generalhealth condition of a subject and severity of a condition being treated.Thus, in some embodiments, a therapeutically effective amount isdetermined empirically. Accordingly, a therapeutically effective amountof a composition that is administered to a subject can be determined byone of ordinary skill in the art based on amounts found effective inanimal or clinical studies, a physician's experience, and suggested doseranges or dosing guidelines, for example.

In certain embodiments, a therapeutically effective amount of acomposition disclosed herein is administered at a suitable dose (e.g.,at a suitable volume, frequency and/or concentration, which oftendepends on a subject's weight, age and/or condition) intended to obtainan acceptable therapeutic outcome. In certain embodiments, atherapeutically effective amount of a composition comprises one or moredoses selected from at least 0.01 mg/kg (e.g., mg of a composition perkg body weight of a subject), at least 0.1 mg/kg, at least 0.5 mg/kg, atleast 1 mg/kg, at least 10 mg/kg or at least 100 mg/kg. In certainembodiments, a therapeutically effective amount of a composition isselected from one or more doses of about 0.001 mg/kg (e.g., mg of acomposition per kg body weight of a subject) to about 5000 mg/kg, 0.01mg/kg to 1000 mg/kg, 0.01 mg/kg to 500 mg/kg, 0.1 mg/kg to 1000 mg/kg, 1mg/kg to 1000 mg/kg, 10 mg/kg to 1000 mg/kg, 100 mg/kg to 1000 mg/kg,0.1 mg/kg to 500 mg/kg, 0.1 mg/kg to 250 mg/kg, 0.1 mg/kg to 150 mg/kg,0.1 mg/kg to 100 mg/kg, 0.1 mg/kg to 75 mg/kg, 0.1 mg/kg to 50 mg/kg,0.1 mg/kg to 25 mg/kg, 0.1 mg/kg to 10 mg/kg, 0.1 mg/kg to 5 mg/kg, 0.5mg/kg to 5 mg/kg, intervening amounts and combinations thereof. In someaspects a therapeutically effective amount of a composition administeredto a subject comprises one or more doses of about 0.1 mg/kg, 0.2 mg/kg,0.3 mg/kg, 0.4 mg/kg, 0.5 mg/kg, 0.6 mg/kg, 0.7 mg/kg, 0.8 mg/kg, 0.9mg/kg, 1 mg/kg, 2 mg/kg, 3 mg/kg, 4 mg/kg, 5 mg/kg, 6 mg/kg, 7 mg/kg, 8mg/kg, 9 mg/kg, 10 mg/kg, 50 mg/kg, 100 mg/kg, 500 mg/kg, andintervening amounts and combinations thereof. In some embodiments atherapeutically effective amount of a composition disclosed herein isbetween about 0.1 mg/kg and about 50 mg/kg.

In certain embodiments, a therapeutically effective amount of acomposition disclosed herein is administered at a suitable volumeintended to obtain an acceptable therapeutic outcome. For example, insome embodiments, a composition herein is administered directly and/ortopically to a damaged tissue. Accordingly, in some embodiments, atherapeutically effective amount of a composition comprises 0.01 ml to50 ml/cm², or about 0.1 ml to 10 ml/cm² (ml of composition/surface areaof damaged tissue).

In some embodiments administering a therapeutically effective amount ofa composition comprises administering a suitable dose at a frequency orinterval as needed to obtain an effective therapeutic outcome. In someembodiments administering a therapeutically effective amount of acomposition disclosed herein comprises administering a suitable dosehourly, every two hours, every 4 hours, every 6 hours, three times aday, twice a day, once a day, six times a week, five times a week, fourtimes a week, three times a week, twice a week, weekly, at combinationsthereof, and/or at regular or irregular intervals thereof, and/or simplyat a frequency or interval as needed or recommended by a medicalprofessional. In some embodiments, a therapeutically effective amount ofa composition is administered continuously by, for example byintravenous administration.

In some embodiments a therapeutically effective amount of a compositionis administered to a subject prior to, during and/or after a subjectreceives a thrombolytic, anti-coagulant or endovascular interventiontherapy. In some embodiments a therapeutically effective amount of acomposition is administered to a subject up to 3 days prior to, up to 2days prior to, up to 1 day prior to, up to 20 hours prior to, up to 15hours prior to, up to 10 hours prior to, up to 5 hours prior to, up to 2hours prior to or up to 1 hour prior to administration of athrombolytic, anti-coagulant or endovascular intervention therapy. Insome embodiments a therapeutically effective amount of a composition isadministered to a subject 0 to 72 hours, 0 and 48 hours, 0 to 24 hours,0 to 12 hours, 0 to 6 hours, 0 to 4 hours, or 0 to 2 hours beforeadministration of a thrombolytic, anti-coagulant or endovascularintervention therapy. In some embodiments a therapeutically effectiveamount of a composition is administered during administration of athrombolytic, anti-coagulant or endovascular intervention therapy. Insome embodiments a therapeutically effective amount of a composition isadministered intermittently or continuously for up to 1 hour after, 2hours after, 4 hours after, 6 hours after, 12 hours after, 24 hoursafter, 2 days after, 3 days after, a week after, 1 month after, 3 monthsafter, 6 months after, 12 months after, 18 months after, 24 months afteror up to 36 months administration of a thrombolytic, anti-coagulant orendovascular intervention therapy.

In some embodiments, a therapeutically effective amount of energy (e.g.,laser energy, light from a light emitting diode, radiofrequency (RF)energy, audio frequency energy, etc.) is administered to a damagedtissue, with or without a composition disclosed herein. In someembodiments, a therapeutically effective amount of energy comprises adose of about 1 Joule/cm² to about 400 Joule/cm², about 1 Joule/cm² toabout 150 Joule/cm², about 1 Joule/cm² to about 100 Joule/cm², about 10Joule/cm² to about 150 Joule/cm², or about 20 Joule/cm² to about 100Joule/cm² (energy of electromagnetic radiation per surface area ofdamaged tissue).

Systems

In some embodiments, one or more present systems may include one or moreof the energy sources and/or devices described herein, one or more ofthe compositions described herein, an imaging device, and/or othercomponents. By way of a non-limiting example, a system may include anLED, a laser, an audio, and/or an RF or audio source (e.g., any one ofthose by itself, any combination of two of those, a combination ofthree, or a combination of all four), and/or one or more (e.g., one,two, three, or more) of the compositions described herein, an imagingdevice, and/or other components.

The present systems may be configured to provide relatively low levelenergy therapy compared to prior systems. The present systems may beconfigured such that therapeutic light (photonic electrons and/or otherelectrons), RF, audio, and/or other electromagnetic energy is absorbedby, but not limited to, chromophores found in units of matter such as,but not limited to atoms, cells, and/or tissue of wounded, cancerous,diseased, aged, aging, damaged, infected, mature, immature, normal,abnormal, undersized, missing and/or contaminated atoms, cells, and/ortissue, to trigger biological reactions that result in beneficialtherapeutic outcomes. In some embodiments, the present systems may beconfigured such that one or more compositions included in a given systemare configured to accept photonic electrons, electrons, and/or otherenergy and to serve as a reservoir of building blocks to assist in newcell organization and to generate new tissue. The present system(s) areconfigured to increase chromophore activity and thus increase cellproliferation, cell migration and generate units of matter such as, butnot limited to tissue. This may assist protein synthesis to regenerateand/or biostimulate units of matter such as, but not limited to tissue,while exhibiting effects such as but not limited to decreasing pain ator surrounding the treatment site and/or decontaminating the tissue ator surrounding the treatment site.

In a natural in vivo state, a chromophore such as but not limited to,cytochrome c oxidase exists within the mitochondria of cells andexhibits chromophore activity which may lead to reactions, such as, butnot limited to biological reactions. Increased chromophore activity isexhibited via utilization of excited photons (photonic electrons) and/orelectrons following their delivery to, but not limited to, chromophoresand other units of matter such as, but not limited to atoms, cells,and/or tissue of wounded, cancerous, diseased, aged, aging, damaged,infected, mature, immature, normal, abnormal, undersized, missing and/orcontaminated atoms, cells, and/or tissue. The utilization of excitedphotons (photonic electrons), other electrons, and/or other energy leadscauses atomic, cellular, and/or tissue generation and/or repair. Theenergy delivered to units of matter such as, but not limited to tissue,is conserved.

The delivery of photons (photonic electrons), electrons, and/or otherenergy and the consequent downstream effect comprises photochemistry andif the energy of the photonic electrons and/or electrons delivered isbio-cell friendly by using specific bio-cell friendly parameters,covalent bonds are not broken and the delivered energy is conservedwithin the biological system. Additionally, if the energy delivered isnot only conserved, but is additionally sufficient for a first excitedsinglet state to be formed, the absorbed electrons can undergointersystem crossing until the electrons reach the long-lived tripletstate within the chromophore. This long life allows an increased amountof reactions to occur once photonic electrons and/or electrons aredelivered into units of matter such as, but not limited to, tissue.

These electron transfer reactions are relevant in the mitochondrialrespiratory chain as follows: the inner mitochondrial membrane containsfive membrane protein complexes: NADH dehydrogenase (Complex I),succinate dehydrogenase (Complex II), cytochrome c reductase (ComplexIII), cytochrome c oxidase (Complex IV), ATP synthase (Complex V), andtwo molecules that freely diffuse, ubiquinone and cytochrome c. Thefreely diffusible molecules shuttle photonic electrons and/or electronsfrom one complex to the next. Insertion of photonic electrons (photons),electrons, and/or other energy into the outer orbital of an atomincreases the shuttling rate of electrons between and among thecomplexes.

For example, the respiratory chain accomplishes the stepwise transfer ofphotonic electrons and/or electrons from NADH and FADH2 (produced in thecitric acid cycle also known as the Krebs cycle) to oxygen molecules toform water molecules. These newly formed water molecules, now harnessingthis electron transfer energy, release this energy and transfer it tothe pumping of protons (H+) from the matrix to the intermembrane space.The gradient of protons formed in the mitochondria across the innermembrane by this process of active transport of photonic electronsand/or electrons forms a miniature battery. Protons are able to flowback down this gradient, re-entering the matrix, through another proteincomplex of the inner membrane, the ATP synthase complex. The exampleshows that energy is unlikely to escape once in the mitochondrialsystem, and is thus conserved for usage by the living system.

Furthermore, the absorption of photonic electrons (photons) and/orelectrons (and/or other radiation and/or energy from an energy sourcedevice as described herein) by molecules such as chromophore moleculesleads to electronically excited states, and consequently leads to anacceleration of electron transfer reactions. The more reduced an initialcell's state, the higher the potential to respond to electron transfer.Notably, cells at the optimal redox state respond weakly or do notrespond.

Units of matter such as, but not limited to atoms, cells, and/or tissueof wounded, cancerous, diseased, aged, aging, damaged, infected, mature,immature, normal, abnormal, undersized, missing and/or contaminatedatoms, cells, and/or tissue having reduced ATP production to no ATPproduction, commonly exist in a reduced, severely reduced or completelyreduced state, thereby significantly exhibit an increased affinity ofvarying degrees of accepting electron transfers. Cytochromes (enzymes)within mitochondria of units of matter such as, but not limited toatoms, cells, and/or tissue of wounded, cancerous, diseased, aged,aging, damaged, infected, mature, immature, normal, abnormal,undersized, missing and/or contaminated atoms, cells, and/or tissue havea high affinity to absorb energy (photonic electrons and/or electrons).Enzymes are catalysts and are able to process thousands of substratemolecules. Energized enzymes, ones that have absorbed electrons, providean amplified tissue response of varying degrees.

By placing at least one composition (substrate) and/or portions thereofinto an excited or soon to be excited mitochondrial “manufacturing”plant, (one that has absorbed or will soon be treated and/or otherwiseirradiated to absorb energy from an energy source device describedherein) the energy-chemical chromophore activity may become furtherincreased and/or accelerated by varying degrees. Providing at least onecomposition and/or portions thereof, and/or energy (e.g., radiation) asdescribed herein, and/or a combination of the two, to units of mattersuch as, but not limited to atoms, cells, and/or tissue of wounded,cancerous, diseased, aged, aging, damaged, infected, mature, immature,normal, abnormal, undersized, missing and/or contaminated atoms, cells,and/or tissue, enables the host units of matter such as, but not limitedto atoms, cells or tissues, to partially or fully rebuild the particularatom, cell and/or tissue and/or tissue segment; depending on the lengthof treatment with the energy and/or the composition used, and/or theoptional method of a treatment cycle.

As described herein, the present systems may be considered relativelylow energy compared to prior systems. If a high energy system is used,the body's circuitry quickly begins to have a backflow of electrons, notallowing further electron deposition. This backflow (back-up) ofelectrons may burn or melt tissue by denaturing proteins. By keeping thesystem a low energy system, as described herein, the system is notlimited to a certain, brief, therapeutic treatment in order to avoidunwanted tissue burning and melting. The therapeutic energy (e.g., at acertain therapeutic threshold) can be maintained for a longer period oftime. Other advantages are contemplated.

The present system(s) are configured for treating units of matter suchas, but not limited to atoms, cells, and/or tissue of wounded,cancerous, diseased, aged, aging, damaged, infected, mature, immature,normal, abnormal, undersized, missing and/or contaminated atoms, cells,and/or tissue, by using one or more energy sources such as but notlimited to a laser photon beam, an LED photon beam, a broadband photonbeam, a radiofrequency electron waveform, an electron current, an audiofrequency electron waveform, and/or at least one composition and/orportions thereof.

The system(s) may be used to deliver photonic electrons and/orelectrons, RF waves, audio waves, and/or other forms of energy intounits of matter such as, but not limited to atoms, cells, and/or tissueof wounded, cancerous, diseased, aged, aging, damaged, infected, mature,immature, normal, abnormal, undersized, missing and/or contaminatedatoms, cells, and/or tissue by using photonic electrons and/or electronssuch as but not limited to a laser photon beam, and/or LED photon beamand/or broadband photon beam and/or radiofrequency electron waveform/sand/or electron current and/or at least one substrate and/or portionsthereof in order to generate units of matter such as, but not limitedto: tissue by using a principle such as, but not limited to,transduction and/or transference.

The system(s) also relate to the novel observation of transfectionand/or transference of units of matter such as, but not limited toelectrons, protons and/or proteins, into tissue that is able to acceptelectrons, protons and/or proteins, such as but not limited to wounded,cancerous, diseased, aged, aging, damaged, infected, mature, immature,normal, abnormal, and/or contaminated units of matter such as, but notlimited to: tissue. Tissue may include vasculature, skin, muscle, nerve,bone origin that may be from ectodermal, endodermal and/or mesodermalorigin, lines and/or pathways, and/or other tissue. The electrons and/orother energy may be generated by an energy source device (e.g., asdescribed herein) configured to generate a laser photon beam, an LEDphoton beam, a broadband photon beam, a radiofrequency waveforms,electron current. The system may be used to add or insert photonicelectrons and/or electrons into tissue that is able to accept electrons,such as but not limited to wounded, cancerous, diseased, aged, aging,damaged, infected, mature, immature, normal, abnormal, and/orcontaminated units of matter such as, but not limited to tissue. In someembodiments, at least one composition and/or portion thereof may beinclude in the system and an associated method (described herein) may beused to generate units of matter such as, but not limited to: tissuethrough a process such as, but not limited to, transfection and/ortransference.

In some embodiments, the system comprises a transfection and/ortransference system wherein a source of energy emits photonic electronsand/or electrons that absorb into outer orbitals of atoms andsubsequently produce ATP, further wherein the source of energyoriginates from one or more of laser photons, LED photons, broadbandphotons, radiofrequency electron waveforms, electron current, and/orother energy sources. Additionally, the present system may include atleast one composition (substrate) and/or portions thereof. The systemmay be used to treat tissue such as, but not limited to wounded,cancerous, diseased, aged, aging, damaged, infected, mature, immature,normal, abnormal, and/or contaminated units of matter such as, but notlimited to tissue. In some embodiments, the presents system isconfigured to facilitate tissue growth of such tissue by delivering theenergy as described. In some embodiments, one or more of the presentsystems may provide for treating deeper and surrounding units of mattersuch as, but not limited to: tissue, by using the principle oftransfection and/or transference.

In some embodiments, one or more of the systems described herein may beuseful for treating tissue, imaging tissue, facilitating tissuegeneration, identifying biomarkers that are common between wound healingand cancer, and/or other uses. For example, a present system can be usedto generate tissue by maintaining a therapeutic energy threshold whiledepositing electrons into outer orbitals, such that ATP is constantlypumped through cells. In the cytoplasm, the ATP is used to assembleproteins. By allowing the system to therapeutically maintain the ATPformation for duration of specific times, the present system is guidesthe body's cells to maintain protein assembly until tissue is formed. Asanother example, the stroma in solid tumors contains a variety ofcellular phenotypes and signaling pathways associated with woundhealing, leading to the concept that a tumor behaves as a wound thatdoes not heal. Similarities between tumors and healing wounds includefibroblast recruitment and activation, extracellular matrix (ECM)component deposition, infiltration of immune cells, neovascularization,and cellular lineage plasticity. However, unlike a wound that heals, theedges of a tumor are constantly expanding(https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6237224/). In someembodiments, a present system may be used to stop the expansion byre-establishing healthy circulation in throughout and surrounding atumor, as the system is configured to do with other tissue. Whencirculation and its balanced vascular endothelial growth factor isre-established, blood vessels grow following a normal course ofarteriogenesis and venogenesis, reestablishing normal patterns ofgrowth.

In some embodiments, one or more of the devices, systems, and/or methodsdescribed herein may be used for lung imaging (e.g., for treatingCovid-19 patients). For example, a Ti-sapphire laser may be configuredto penetrate energy through skin to the lung tissue. Reflected,transmitted, or scattered light can be captured by a photomultipliertube (PMT) followed by a high dynamic range oscilloscope. A laser mayhave a sufficient diameter for treatment of lungs to treat a twocentimeter cubed area or more, a ten centimeter cubed area or more, oran entire lung or lungs, for example. In some embodiments, these areasmay be covered by more than one laser. Such a laser (or lasers) may havea custom housing unit sufficient enough in size for one or more lasers.The imaging devices and/or systems may include a time stretch dispersiveFourier transform system for real-time spectral measurement. Depth ofpenetration, reflection, transmission, or scattering on lung tissue maybe determined using PMT, a high dynamic range oscilloscope and a timestretch dispersive Fourier transform system, for example. In someembodiments, the imaging devices and/or systems may include an opticalcoherence tomography system. Software may provide for 3D volumetricvisualization.

Methods

In some embodiments, a method comprises treating a wound or damagedtissue of a subject, which method comprises administering atherapeutically effective amount of a composition disclosed herein tothe wound or damaged tissue, and/or administering a therapeuticallyeffective amount of energy (e.g., laser energy, light from a lightemitting diode, radiofrequency (RF) energy, audio frequency energy,etc.), as described herein, to the wound or damaged tissue. In someembodiments, the energy (e.g., laser energy, light from a light emittingdiode, radiofrequency (RF) energy, audio frequency energy, etc.) isadministered before, during and/or after administration of thecomposition to the wound or damaged tissue. In some embodiments, acomposition is administered topically or locally. In some embodiments, acomposition is administered orally.

In some embodiments, the energy (e.g., laser energy, light from a lightemitting diode, radiofrequency (RF) energy, audio frequency energy,etc.) administered comprises a wavelength in a range of 200 nm to 2500nm. In some embodiments, the energy administered comprises a wavelengthin a range of 700-900 nm. In some embodiments, the energy administeredcomprises a wavelength in the RF range, for example a frequency in arange of between about 1 dm and 1 km. In some embodiments, atherapeutically effective amount energy administered comprises a dose of1 to 400 joules/cm² or 10 to 100 joules/cm². In some embodiments, atherapeutically effective amount of energy that is administered to awound or damaged tissue is administered for about 0.1 to 30 minutes. Insome embodiments, a therapeutically effective amount of energy that isadministered to a wound or damaged tissue is administered for about 1 to30 min., 1 to 15 min., or for about 1 to 5 min. In some embodiments, atherapeutically effective amount of energy that is administered to awound or damaged tissue comprises a dose of 10 to 100 joules/cm²administered for about 1 to 15 minutes at a wavelength in a range of700-900 nm. A treatment may comprise administering one, or multipledoses of energy.

In some embodiments, a method comprises administering a therapeuticallyeffective amount of the composition by local or systemic administration.In certain embodiments, a therapeutically effective amount of thecomposition is administered topically or directly to the wound ordamaged tissue. In certain embodiments, a therapeutically effectiveamount of a composition comprises a volume in a range of 0.01 to 20ml/cm². Accordingly, in some embodiments, a wound or damaged tissue istreated by administering 0.01 to 20 ml/cm² of a composition directly tothe wound or damaged tissue and administering a therapeutic amount ofenergy (e.g., laser energy, light from a light emitting diode,radiofrequency (RF) energy, audio frequency energy, etc.) to the woundor damaged tissue that comprises the composition, or a portion thereof.

In certain embodiments, a therapeutically effective amount of acomposition comprises a dose of about 0.1 mg/cm² to 10 gram/cm² (weightof composition/surface area of damaged tissue). Accordingly, in someembodiments, a wound or damaged tissue is treated by administering 0.1mg/cm² to 10 gram/cm² of a composition directly to the wound or damagedtissue and administering a therapeutic amount of energy (e.g., laserenergy, light from a light emitting diode, radiofrequency (RF) energy,audio frequency energy, etc.) to the wound or damaged tissue thatcomprises the composition, or a portion thereof. The energy acts as adelivery system of the composition and when treatment repeated, anotherdose is dispensed to be used on a wound or damaged tissue.

In certain embodiments, a therapeutically effective amount of acomposition is administered systemically (e.g., orally or by parenteraladministration) at a dose of 0.1 to 50 mg/kg (weight of composition/bodyweight of the subject). Accordingly, in some embodiments, a wound ordamaged tissue is treated by administering a dose of 0.1 to 50 mg/kg ofa composition to a subject by systemic administration followed byadministering a therapeutic amount of energy (e.g., laser energy, lightfrom a light emitting diode, radiofrequency (RF) energy, audio frequencyenergy, etc.) to the wound or damaged tissue.

In certain embodiments, a therapeutically effective amount of acomposition is administered as a mouth wash or oral rinse, for examplefor treatment of damaged tissue located on the tongue or in the mouth.Accordingly, in some embodiments, 5-100 ml of a composition describedherein is introduced into the mouth of a subject such that thecomposition contacts a damaged tissue within the mouth for a period of0.1 to 3 minutes, followed by the subject expelling or spitting thecomposition from the mouth. After administering the composition, thewound or damaged tissue is treated by administering a therapeutic amountof energy (e.g., laser energy, light from a light emitting diode,radiofrequency (RF) energy, audio frequency energy, etc.) to the woundor damaged tissue.

A therapeutically effective amount of energy, in some embodiments, isadministered prior to administration of a composition described herein.Accordingly, a therapeutically effective amount of energy, in someembodiments, is administered 0.1 min. to 1 hour before administration ofa composition described herein. In some embodiments, a therapeuticallyeffective amount of energy is administered 1 min. to 30 min., or 1 min.to 15 min. before administration of a composition described herein.

Kits

In some embodiments, provided herein is a kit comprising a compositiondisclosed herein or a pharmaceutical composition comprising acomposition disclosed herein. In some embodiments, a kit comprises oneor more doses of a pharmaceutical composition comprising a compositiondisclosed herein. In some embodiments, a kit comprises one or more packsand/or one or more dispensing devices, which can contain one or moredoses of a composition disclosed herein, or pharmaceutical compositionthereof, as described herein. Non-limiting examples of a pack include ametal, glass, or plastic container, syringe or blister pack thatcomprises a composition disclosed herein or a composition describedherein. In certain embodiments, a kit comprises a dispensing device suchas a syringe or inhaler, that may or may not comprise a compositiondisclosed herein or a composition described herein. A pack and/ordispenser device can be accompanied by instructions for administration.The pack or dispenser can also be accompanied with a notice associatedwith the container in a form prescribed by a governmental agencyregulating the manufacture, use, or sale of pharmaceuticals, whichnotice is reflective of approval by the agency of the form of the drugfor human or veterinary administration. Such notice, for example, can bethe labeling approved by the U.S. Food and Drug Administration forprescription drugs, or the approved product insert.

In some embodiments a kit or pack comprises an amount of a compositiondisclosed herein sufficient to treat a patient for 1 day to 1 year, 1day to 180 days, 1 day to 120 days, 1 day to 90 days, 1 day to 60 days,1 day to 30 days, 1-24 hours, 1-12 hours, 1-4 hours, or amount of timethere between.

A kit optionally includes a product label and/or one or more packaginginserts, that provide a description of the components or instructionsfor use in vitro, in vivo, or ex vivo, of the components therein.Exemplary instructions may include instructions for a treatment protocolor therapeutic regimen. In certain embodiments, a kit comprisespackaging material, which refers to a physical structure housingcomponents of the kit. The packaging material can maintain thecomponents sterilely and can be made of material commonly used for suchpurposes (e.g., paper, corrugated fiber, glass, plastic, foil, ampules,vials, tubes, etc.). Product labels or inserts include “printed matter,”e.g., paper or cardboard, or separate or affixed to a component, a kitor packing material (e.g., a box), or attached to an ampule, tube orvial containing a kit component. Labels or inserts can additionallyinclude a computer readable medium, optical disk such as CD- orDVD-ROM/RAM, DVD, MP3, magnetic tape, or an electrical storage mediasuch as RAM and ROM or hybrids of these such as magnetic/optical storagemedia, FLASH media or memory-type cards. Product labels or inserts caninclude identifying information of one or more components therein, doseamounts, clinical pharmacology of the active ingredient(s) includingmechanism of action, pharmacokinetics (PK) and pharmacodynamics (PD).Product labels or inserts can include information identifyingmanufacturer information, lot numbers, manufacturer location, date,information on an indicated condition, disorder, disease or symptom forwhich a kit component may be used. Product labels or inserts can includeinstructions for the clinician or for a subject for using one or more ofthe kit components in a method, treatment protocol or therapeuticregimen. Instructions can include dosage amounts, frequency or duration,and instructions for practicing any of the methods, treatment protocolsor therapeutic regimes set forth herein. A kit can additionally includelabels or instructions for practicing any of the methods describedherein. Product labels or inserts can include information on potentialadverse side effects and/or warnings.

EXAMPLES Example 1—An Exemplary Fibrous Collagen Composition

Ingredients:

-   -   a) 1 g dried collagen fibers    -   b) 2.3 g HCl    -   c) 0.5 g hyaluronic acid (HA) powder    -   d) 0.1 gCu or 0.15 gCuCl₂ or 0.15 gCuCl₃    -   e) 0.1 g Fe or 0.15 g FeCl₃

Collagen fiber (1 g) was added to a 1N HCl solution (2.3 g) andincubated for 14-18 hours with intermittent stirring. The insolublecollagen was pelleted by centrifugation and the supernatant was removed.The damp pellet was resuspended in isopropanol (99-100%) for 60 minutesat 37° C. with intermittent stirring. The partially hydrolyzed collagenwas fluffed dry by separating the fiber clumps as much as possibleduring drying. Drying was finished in a fluidized bed at 45° C.Hyaluronic acid was added and mixed with a vortexer. The iron, or ironsalt, was added and mixed, then the copper or copper salt was added andmixed again with a vortexer. The final pH of the dry composition wasabout 2-3 as measured by litmus paper (pH 0-14 LAB RAM SUPPLIES pHLitmus paper). Additional ingredients can be added as needed to completeeach of formulas 1-23 as shown in the tables of FIGS. 35-47. Metals wereadded at 0.1 g per composition, metal salts at 0.15 g per compositionand other ingredients at 0.5 g per composition.

Example 2—An Exemplary Powdered Collagen Composition

Ingredients:

-   -   a) 1 g dried collagen fibers    -   b) 2.3 g HCl    -   c) 0.5 g hyaluronic acid (HA) powder    -   d) 0.1 g Cu or 0.15 g CuCl₂ or 0.15 g CuCl₃    -   e) 0.1 g Fe or 0.15 g FeCl₃

Collagen fiber (1 g) was added to a 1N HCl solution (2.3 g) andincubated for 14-18 hours with intermittent stirring. The insolublecollagen was pelleted by centrifugation and the supernatant was removed.The damp pellet was resuspended in isopropanol (99-100%) for 60 minutesat 37° C. with intermittent stirring. The partially hydrolyzed collagenwas fluffed dry by separating the fiber clumps as much as possibleduring drying. Drying was finished in a fluidized bed at 45° C. Thedried fiber was milled to specification (e.g., 20 mesh size fibersinstead of fiber clumps) using a 20-mesh screen and an 18-gage needle.

Hyaluronic acid was added to the milled collagen and mixed with avortexer. The iron, or iron salt, was added and mixed, then the copperor copper salt was added and mixed again with a vortexer. The final pHof the dry composition was about 2-3 as measured by litmus paper (pH0-14 LAB RAM SUPPLIES pH Litmus paper). Additional ingredients can beadded as needed to complete each of formulas 1-23 as shown in the tablesof FIGS. 35-47. Metals were added at 0.1 g per composition, metal saltsat 0.15 g per composition and other ingredients at 0.5 g percomposition.

Example 3—An Exemplary HA Gel Composition

Ingredients:

-   -   a) 30 g HA powder    -   b) 5 g Collagen    -   c) 0.01 g-0.3 g HCl    -   d) 473.18 ml water    -   e) 0.3 g Fe or 0.15 g FeCl₁₃    -   f) 0.3 g Cu or 0.15 g CuCl₂ or 0.15 g CuCl₃

All ingredients were added to a laboratory blending dish and mixed. Thefinal pH of the dry composition was about 3-7 as measured by litmuspaper (pH 0-14 LAB RAM SUPPLIES pH Litmus paper). Additional ingredientscan be added as needed to complete each of formulas 1-23 as shown in thetables of FIGS. 35-47. Metals were added at 0.3 g per composition, metalsalts at 0.15 g per composition and other ingredients at 0.5 g percomposition.

Example 4—Treatment of Acute Wound (Tooth Extraction Site

A tip of a laser energy source as described herein was inserted onto thegingiva, just above the periodontal ligament (pdl) of a tooth prior toextraction (1 mm distance from the light source to the pdl). The laserwas set at a wavelength of 850 nm. The area was treated by moving thelaser circumferentially for 1 minute. The tooth was extracted leaving anextraction site wound of about 4-5 mm in diameter. The extraction sitewas treated for 1 minute by placing the Evo-lase tip on the extractionsite and rotating for 1 minute. A composition (substrate; 0.6 ml) asindicated in the table of FIG. 31 (substrate and laser treatment),formulated as a fibrous mass (Example 1), was placed into the extractionsite while disrupting blood clotting. After 1 minute, the extractionsite was treated by placing the laser tip over the extraction site (1 mmdistance from energy source to extraction site) and rotating for 2minutes taking care not to disrupt blood clotting. The total treatmenttime was 4 minutes at 13.2 Joules/cm² at a wavelength of 850 nm. Thecompositions and data for tooth extraction sites treated with substratealone (0.6 ml; no laser treatment), or laser alone are shown in thetable of FIGS. 35 & 39, respectively.

Example 5—Treatment of Acute Wound (Head

A tip of a laser energy source as described herein was placed onto anacute wound of the head (1 mm distance from the light source to thewound) having a diameter of about 3-5 mm. The laser was set at awavelength of 850 nm. The area was treated by moving the lasercircumferentially for 1 minute. A composition (substrate; 0.3-0.6 ml) asindicated in the table of FIG. 41, Formulation 3, formulated as afibrous mass (Example 1), was placed onto the wound while disruptingblood clotting. The wound was immediately covered with clear tape. After1 minute, the wound site was treated by placing the energy source tipover the wound site (1 mm distance from energy source to wound site) androtating for 2 minutes. The total treatment time was 3 minutes at 9.9Joules/cm² at a wavelength of 850 nm. The tape was left on until thetissue has grown epithelium. The data for head wound sites treated withsubstrate alone (0.3-0.6 ml; no laser treatment), or laser andsubstrate, are shown in the tables of FIGS. 41 and 40 (Formulation 3),respectively. FIG. 41 shows a table of results for the treatment ofacute wounds with the substrates of the indicated formulas alone. Therow of Formulation #3 provides data for a head wound.

Example 6—Treatment of Acute Wound (Leg)

A tip of a laser energy source as described herein was placed onto anacute wound of the leg (1 mm distance from the wound) having dimensionsof about 4 cm×6 cm. The laser was set at a wavelength of 850 nm. Thewound area was treated by moving the laser circumferentially for 7minutes. A composition (substrate; 10 ml) as indicated in table of FIG.38 (substrate and laser treatment), formulated as a gel, was placed ontothe wound. The wound was immediately covered with clear tape. The woundsite was treated by placing the tip over the wound site (1 mm distancefrom energy source to wound site) and rotating for 7 minutes. The totaltreatment time was 15 minutes at 50 Joules/cm² at a wavelength of 850nm. The tape was left on until the next appointment. The data for legwound sites treated with substrate alone (10 ml; no laser treatment), orlaser alone, are shown in table of FIGS. 37 and 39, respectively.

Example 7—Treatment of Diabetic Foot

A tip of a laser energy source as described herein was placed over adark blue area of a diabetic foot lacking circulation (3 mm distancefrom the affected area). The laser was set at a wavelength of 850 nm.The affected area was treated for 15 minutes. A composition (substrate;10 ml) as indicated in the table of FIG. 43 (substrate and lasertreatment), formulated as a gel, was placed onto the affected area untilthe gel absorbed into the tissue. The affected area was again treatedwith the energy source (3 mm distance from energy source to affectedarea) for 15 minutes. The total treatment time was 30 minutes at 100Joules/cm² at a wavelength of 850 nm. The data for treatment withsubstrate alone (10 ml; no laser treatment), or laser alone, are shownin the tables of FIGS. 38 (formulation 20) and 39 (formulation 20),respectively. Circulation improvement was detected by a change in colorof the affected area from dark blue to pinkish/red after 3 weeks ofdaily self-administered treatment and the patient described the affectedarea as feeling warm after treatment.

Example 8—Treatment of Obstructed Circulation

A tip of a laser energy source as described herein was placed over adark blue area of a leg (1 cm in diameter) having circular obstruction(1 mm distance from the affected area to the light source). The laserwas set at a wavelength of 810 nm. The affected area was treated for 15minutes. A composition (substrate; 5 ml) as indicated in the table ofFIG. 43 (Formulation 20) (substrate and laser treatment), formulated asa gel, was placed onto the affected area until the gel absorbs into thetissue. The affected area was again treated with the energy source (1 mmdistance from energy source to affected area) for 15 minutes. The totaltreatment time was 30 minutes at 36 Joules/cm² at a wavelength of 810nm. The data for treatment with substrate alone (5 ml; no lasertreatment), or laser alone, are shown in the table of FIGS. 41(formulation 20) and 42 (formulation 20), respectively. Circulationimprovement was observed by a change in color of the affected area asthe dark blue 1 cm diameter obstruction diminished in color. After 5treatments, the dark blue obstruction was observed to have disappearedand the tone of the tissue was an even warm pinkish/olive color.

Example 9—Treatment of Tongue and/or Surrounding Tissue

-   -   (1) Drink 4 oz of diluted Substrate 1, 2, 3, 4, 5 and/or 6.    -   (2) Wait 15 minutes.    -   (3) Optionally repeat 1 and/or 2.    -   (4) Direct RF/laser, RF or laser energy at tongue and/or        surrounding tissue of the oral cavity and head/neck region. Keep        energy in place for 10-20 minutes or until desired effect is        achieved.    -   (5) Repeat step 4 until desired result is achieved.    -   (6) Procedure can start at step 1 or step 4.        Non-limiting examples of surrounding tissues include bone,        cartilage, muscles, tendons, nerves, blood vessels, epithelium,        the like or combinations thereof.

Example 10—Certain Non-Limiting Embodiments

A1. A system for use in tissue generation, comprising:

-   -   (a) providing at least one device comprising a light, radio        frequency and/or electric current energy source with at least        one opening and/or optionally with at least one opening of a        handpiece, fiber, panel, pad, lead and/or current through which        the light, radio frequency and/or electric current energy source        can emit a beam of light and/or a radio frequency waveform        and/or an electric current, further wherein the device is        connected to a power source;    -   (b) providing at least one substrate and/or portion/s thereof,        wherein the at least one substrate and/or portion/s thereof is        applied to a tissue;    -   (c) contacting the tissue with the beam of light and/or the        radio frequency waveform and/or the electron current via the        handpiece,    -   wherein the contacting of the tissue occurs before, during or        after the at least one substrate and/or portion/s thereof is        applied to the tissue, further wherein the at least one device        and the at least one substrate and/or portion/s thereof is        capable of inducing a biological reaction.        A2. A system for use in repairing tissue subsequent to an acute        or chronic injury to an individual, comprising:    -   (a) a device comprising a light, radio frequency and/or electric        current energy source with at least one opening, through which        the light, radio frequency or electric current energy source can        emit a beam of light and/or a radio frequency waveform and/or an        electric current, further wherein the instrument is connected to        a power source;    -   (b) at least at least one substrate and/or portion/s thereof,        wherein the at least one substrate and/or portion/s thereof is        applied to the tissue;    -   (c) at least at least one substrate and/or portion/s thereof,        wherein the at least one substrate and/or portion/s thereof is        applied to the tissue, further wherein optionally the beam of        light and/or the radio frequency waveform and/or the electron        current is placed over the tissue;    -   (d) at least one energy source wherein the beam of light and/or        the radio frequency waveform and/or the electron current is        placed over the tissue, further wherein optionally the at least        one substrate and/or portion/s thereof is applied to the tissue.

The entirety of each patent, patent application, publication or anyother reference or document cited herein hereby is incorporated byreference. In case of conflict, the specification, includingdefinitions, will control.

Citation of any patent, patent application, publication or any otherdocument is not an admission that any of the foregoing is pertinentprior art, nor does it constitute any admission as to the contents ordate of these publications or documents.

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Although methods and materialssimilar or equivalent to those described herein can be used in thepractice or testing of the present invention, suitable methods andmaterials are described herein.

All of the features disclosed herein may be combined in any combination.Each feature disclosed in the specification may be replaced by analternative feature serving a same, equivalent, or similar purpose.Thus, unless expressly stated otherwise, disclosed features (e.g.,antibodies) are an example of a genus of equivalent or similar features.

The phrase “induced by”, encompasses “worsened by”, “aggravated by”,“exacerbated by”, and/or “magnified by”, unless clearly indicatedotherwise.

As used herein, all numerical values or numerical ranges includeintegers within such ranges and fractions of the values or the integerswithin ranges unless the context clearly indicates otherwise. Further,when a listing of values is described herein (e.g., about 50%, 60%, 70%,80%, 85% or 86%) the listing includes all intermediate and fractionalvalues thereof (e.g., 54%, 85.4%). Thus, to illustrate, reference to 80%or more identity, includes 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%,90%, 91%, 92%, 93%, 94% etc., as well as 81.1%, 81.2%, 81.3%, 81.4%,81.5%, etc., 82.1%, 82.2%, 82.3%, 82.4%, 82.5%, etc., and so forth.

Reference to an integer with more (greater) or less than includes anynumber greater or less than the reference number, respectively. Thus,for example, a reference to less than 100, includes 99, 98, 97, etc. allthe way down to the number one (1); and less than 10, includes 9, 8, 7,etc. all the way down to the number one (1).

As used herein, all numerical values or ranges include fractions of thevalues and integers within such ranges and fractions of the integerswithin such ranges unless the context clearly indicates otherwise. Thus,to illustrate, reference to a numerical range, such as 1-10 includes 1,2, 3, 4, 5, 6, 7, 8, 9, 10, as well as 1.1, 1.2, 1.3, 1.4, 1.5, etc.,and so forth. Reference to a range of 1-50 therefore includes 1, 2, 3,4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, etc., upto and including 50, as well as 1.1, 1.2, 1.3, 1.4, 1.5, etc., 2.1, 2.2,2.3, 2.4, 2.5, etc., and so forth.

Reference to a series of ranges includes ranges which combine the valuesof the boundaries of different ranges within the series. Thus, toillustrate reference to a series of ranges, for example, of 1-10, 10-20,20-30, 30-40, 40-50, 50-60, 60-75, 75-100, 100-150, 150-200, 200-250,250-300, 300-400, 400-500, 500-750, 750-1,000, 1,000-1,500, 1,500-2,000,2,000-2,500, 2,500-3,000, 3,000-3,500, 3,500-4,000, 4,000-4,500,4,500-5,000, 5,500-6,000, 6,000-7,000, 7,000-8,000, or 8,000-9,000,includes ranges of 10-50, 50-100, 100-1,000, 1,000-3,000, 2,000-4,000,etc.

Modifications can be made to the foregoing without departing from thebasic aspects of the technology. Although the technology has beendescribed in substantial detail with reference to one or more specificembodiments, those of ordinary skill in the art will recognize thatchanges can be made to the embodiments specifically disclosed in thisapplication, yet these modifications and improvements are within thescope and spirit of the technology.

The invention is generally disclosed herein using affirmative languageto describe the numerous embodiments and aspects. The invention alsospecifically includes embodiments in which particular subject matter isexcluded, in full or in part, such as substances or materials, methodsteps and conditions, protocols, or procedures. For example, in someembodiments or aspects of the methods disclosed herein, some materialsand/or method steps are excluded. Thus, even though the invention isgenerally not expressed herein in terms of what the invention does notinclude aspects that are not expressly excluded in the invention arenevertheless disclosed herein.

Some embodiments of the technology described herein suitably can bepracticed in the absence of an element not specifically disclosedherein. Accordingly, in some embodiments the term “comprising” or“comprises” can be replaced with “consisting essentially of” or“consisting of” or grammatical variations thereof. The term “a” or “an”can refer to one of or a plurality of the elements it modifies (e.g., “areagent” can mean one or more reagents) unless it is contextually cleareither one of the elements or more than one of the elements isdescribed. The term “about” as used herein refers to a value within 10%of the underlying parameter (i.e., plus or minus 10%), and use of theterm “about” at the beginning of a string of values modifies each of thevalues (i.e., “about 1, 2 and 3” refers to about 1, about 2 and about3). For example, a weight of “about 100 grams” can include weightsbetween 90 grams and 110 grams. The term “substantially” as used hereinrefers to a value modifier meaning “at least 95%”, “at least 96%”, “atleast 97%”, “at least 98%”, or “at least 99%” and may include 100%. Forexample, a composition that is substantially free of X, may include lessthan 5%, less than 4%, less than 3%, less than 2%, or less than 1% of X,and/or X may be absent or undetectable in the composition.

Further embodiments are described in the following list of clauses:

-   1. A composition with wound healing and tissue regenerative    properties, the composition comprising two or more of, or three or    more of: collagen, hyaluronic acid, fucose, copper, and iron.-   2. The composition of clause 1, wherein the collagen comprises    collagen fibers.-   3. The composition of clause 1 or 2, wherein the collagen comprises    partially or completely hydrolyzed collagen.-   4. The composition of any one of clauses 1 to 3, wherein the    collagen is acid treated collagen.-   5. The composition of any one of clauses 1 to 4, wherein the copper    comprises a copper salt.-   6. The composition of clause 5, wherein the copper salt comprises    CuCl₂ or CuCl₃.-   7. The composition of any one of clauses 1 to 6, wherein the iron    comprises an iron salt.-   8. The composition of clause 7, wherein the iron salt comprises    FeCl₃.-   9. The composition of any one of clause 1 to 8, wherein the    composition has a pH in a range of 1 to 4, or 2 to 3.-   10. The composition of any one of clauses 1 to 9, wherein the    composition comprises a water content in a range of 0% to 20%, or 0%    to 10%, or 0% to 5% (wt/wt).-   11. The composition of any one of clauses 1 to 10, wherein the    composition is in the form of a powder, foam, paste, or gel.-   12. The composition of any one of clauses 1 to 11, where the    percentage of collagen in the composition is a range of 25-70%,    40-60% or 50-60% (wt/wt).-   13. The composition of any one of clauses 1 to 11, wherein the    percentage of collagen in the composition is about 55-60% (wt/wt).-   14. The composition of any one of clauses 1 to 13, wherein the    percentage of hyaluronic acid in the composition is in a range of    20-80%, 25-35% or 25-30% (wt/wt).-   15. The composition of any one of clauses 1 to 14, wherein the    percentage of copper and/or iron in the composition is in a range of    0.1-15%, 1-15%, or 5%-10% (wt/wt).-   16. The composition of any one of clauses 1 to 15, wherein the    collagen is present in the composition in a range of 50-70% (wt/wt),    the hyaluronic acid is present in the composition in a range of    25-35% (wt/wt), the copper is present in the composition in a range    of 4-10% (wt/wt) and the iron is present in the composition in a    range of 4-10% (wt/wt).-   17. The composition of any one of clauses 1 to 16, wherein the    composition comprises HCl.-   18. The composition of any one of clauses 1 to 17, wherein the    percentage of HCL in the composition is in a range of 0.0005-0.5%    (wt/wt).-   19. The composition of any one of clauses 1 to 8, wherein the    composition has a pH in a range of 2 to 3, or 3 to 7.-   20. The composition of any one of clauses 1 to 8 and 19, wherein the    composition comprises a water content in a range of 75% to 99%, or    85% to 99%, or 90% to 95% (wt/wt).-   21. The composition of any one of clause 1 to 8 and 19 to 20,    wherein the composition is in the form of a gel, emulsion, slurry or    a liquid.-   22. The composition of any one of clauses 1 to 8 and 19 to 21,    wherein the percentage of collagen in the composition is a range of    0.005-5% or 0.01-1% (wt/wt).-   23. The composition of any one of clauses 1 to 8 and 19 to 22,    wherein the percentage of collagen in the composition is about 0.1%    (wt/wt).-   24. The composition of any one of clauses 1 to 8 and 19 to 23,    wherein the percentage of hyaluronic acid in the composition is in a    range of 0.01-10%, 0.05-5%, or 0.1-1% (wt/wt).-   25. The composition of any one of clauses 1 to 8 and 19 to 24,    wherein the percentage of copper and/or iron in the composition is    in a range of 0.001%-5%, 0.005%-1% or 0.01%-0.1% (wt/wt).-   26. The composition of any one of clauses 1 to 8 and 19 to 25,    wherein the collagen is present in the composition in a range of    0.01-0.5% (wt/wt), the hyaluronic acid is present in the composition    in a range of 0.1-1.5% (wt/wt), the copper is present in the    composition in a range of 0.01%-0.1% (wt/wt) and the iron is present    in the composition in a range of 0.01%-0.1% (wt/wt).-   27. The composition of any one of clauses 1 to 8 and 19 to 26,    wherein the composition comprises HCl.-   28. The composition of any one of clauses 1 to 8 and 19 to 27,    wherein the percentage of HCL in the composition is in a range of    0.0005-0.5% (wt/wt).-   29. The composition of any one of clauses 1 to 28, further    comprising gold or a salt thereof.-   30. The composition of clause 29, wherein the gold comprises gold    particles or gold nanoparticles.-   31. The composition of clause 29 or 30, wherein the percentage of    gold in the composition is in a range of 0.001%-10%, 0.001%-5%, or    0.01%-0.1% (wt/wt).-   32. The composition of any one of clauses 1 to 31, further    comprising silver or a salt thereof.-   33. The composition of clause 32, wherein the percentage of silver    in the composition is in a range of 0.001%-10%, 0.001%-5%, or    0.01%-0.1% (wt/wt).-   34. The composition of any one of clauses 1 to 33, further    comprising Fucose.-   35. The composition of clause 34, wherein the percentage of Fucose    in the composition is in a range of 0.01%-30%, 0.05%-15%, 0.05% to    2% or 20-25% (wt/wt).-   36. The composition of any one of clauses 1 to 35, further    comprising adenosine triphosphate.-   37. The composition of clause 36, wherein the percentage of    adenosine triphosphate in the composition is in a range of    0.01%-30%, 0.05%-15%, 0.05% to 2% or 20-25% (wt/wt).-   38. The composition of any one of clauses 1 to 37, further    comprising acetylcholine.-   39. The composition of clause 38, wherein the percentage of    acetylcholine in the composition is in a range of 0.01%-30%,    0.05%-15%, 0.05% to 2% or 20-25% (wt/wt).-   40. The composition of any one of clauses 1 to 39 wherein the    composition comprises collagen and copper.-   41. The composition of any one of clauses 1 to 40 wherein the    composition comprises collagen and iron.-   42. The composition of any one of clauses 1 to 41 wherein the    composition comprises collagen, copper and iron.-   43. The composition of any one of clauses 1 to 42, wherein the    composition comprises hyaluronic acid and copper.-   44. The composition of any one of clauses 1 to 43, wherein the    composition comprises hyaluronic acid and iron.-   45. The composition of any one of clauses 1 to 44, wherein the    composition comprises hyaluronic acid, iron and copper.-   46. The composition of any one of clauses 1 to 45 wherein the    composition comprises hyaluronic acid and fucose.-   47. The composition of any one of clauses 1 to 46, further    comprising fucose and/or silver.-   48. The composition of any one of clauses 1 to 47, further    comprising gold.-   49. The composition of any one of clauses 1 to 47, further    comprising ATP and/or acetylcholine.-   50. The composition of any one of clauses 1 to 49, wherein the    composition is formulated as a pharmaceutical composition.-   51. The composition of any one of clauses 1 to 50, wherein the    pharmaceutical composition is formulated as a tablet, capsule,    emulsion, powder, gel, paste, liquid, or spray,-   52. The composition of any one of clauses 1 to 51, wherein the    pharmaceutical composition is formulated for delivery by an inhaler.-   53. A kit comprising the composition or pharmaceutical composition    of any one of clauses 1 to 52.-   54. The composition, pharmaceutical composition or kit of any one of    clauses 1 to 53 for use in treating a wound in a subject, inducing    tissue regeneration in a subject, or improving circulation in a    tissue of a subject.-   55. A method of treating a wound or tissue damage in a subject    comprising administering a therapeutically effective amount of the    composition of any one of clauses 1 to 52 to the subject.-   56. A method of inducing tissue regeneration in a subject comprising    administering a therapeutically effective amount of the composition    of any one of clauses 1 to 52 to a wound of the subject.-   57. A method of improving circulation in a subject comprising    administering a therapeutically effective amount of the composition    of any one of clauses 1 to 52 to a tissue of the subject.-   58. The method of any one of clauses 55 to 57, further comprising    administering a therapeutically effective amount of energy to the    wound or damaged tissue of the subject.-   59. The method of clause 58, wherein the energy comprises a    wavelength in a range of 350 nm to 1400 nm.-   60. The method of clause 59, wherein the energy comprises a    wavelength in the range of 700-900 nm.-   61. The method of any one of clauses 58 to 60, wherein the    therapeutically effective amount of energy comprises a dose of 1 to    400 joules/cm² or 10 to 100 joules/cm².-   62. The method of clause 61, wherein the dose is delivered to the    wound or tissue for 1 to 30 minutes.-   63. The method of any one of clauses 58 to 62, wherein the    therapeutically effective amount of energy is administered 1 to 20    times.-   64. The method of any one of clauses 58 to 63, wherein the energy is    administered before and/or after administration of the composition.-   65. The method of any one of clauses 58 to 64, wherein the    composition is administered by local or systemic administration; by    topical, oral, sublingual, parenteral, intranasal or intratracheal    administration; or by inhalation.-   66. The method of any one of clauses 58 to 65, wherein the    therapeutically effective amount of the composition comprises a    volume in a range of 0.1 to 20 ml, or 0.1 to 10 ml.-   67. The method of any one of clauses 58 to 66, wherein the    therapeutically effective amount of the composition comprises a dose    of 0.1 to 50 mg/kg of body weight of the subject when administered    systemically.-   68. The method of any one of clauses 58 to 67, wherein the    therapeutically effective amount of the composition comprises a dose    of 0.1 mg/cm² to 1 gram/cm² when administered locally or topically.-   69. The method of any one of clauses 58 to 68, wherein the wound is    an acute wound or a chronic wound.-   70. The method of clause 69, wherein the acute wound is a surgical    incision.-   71. The method of clause 69, wherein the acute wound is a tooth    extraction site.-   72. The method of clause 69, wherein the chronic wound is an ulcer.-   73. The method of any one of clauses 58 to 68, wherein the tissue is    hypoxic, ischemic, lacks sufficient circulation, is restricted of    blood flow and/or comprises a vasculature obstruction.-   74. The method of any one of clauses 58 to 68, wherein the subject    is diabetic.-   75. A system configured to be used to treat and/or generate tissue,    the system comprising:    -   a device configured to generate one or more types of therapeutic        energy for provision to the tissue, the one or more types of        therapeutic energy comprising one or more of laser radiation,        radio frequency (RF) waves, audio frequency waves, or light from        a light emitting diode (LED); and    -   a composition with wound healing and/or tissue regenerative        properties, the composition comprising two or more of, or three        or more of: collagen, hyaluronic acid, fucose, copper, and iron.-   76. The system of clause 75, wherein the collagen is present in the    composition in a range of 0.01-0.5% (wt/wt), the hyaluronic acid is    present in the composition in a range of 0.1-1.5% (wt/wt), the    copper is present in the composition in a range of 0.01%-0.1%    (wt/wt) and the iron is present in the composition in a range of    0.01%-0.1% (wt/wt).-   77. The system of clause 75, wherein the device comprises a laser    configured to generate the laser radiation, the laser comprising one    or more individual fibers configured to conduct the laser radiation,    the laser radiation having a power of about 0.1-5 W per fiber.-   78. The system of clause 75, wherein the one or more types of    therapeutic energy have a wavelength of about 200 nm to about 2500    nm.-   79. The system of clause 75, wherein the device comprises an LED    configured to generate light comprising photons configured to be    received by the tissue, the LED arranged on a board with a plurality    of other LEDs configured to generate the same and/or similar light.-   80. The system of clause 75, wherein the device comprises an RF    and/or an audio frequency function generator, the RF and/or audio    frequency comprising a frequency of about 100 Hz-4 MHz.-   81. A device configured to generate one or more types of therapeutic    energy for provision to tissue, the device comprising:    -   one or more energy sources configured to generate one or more        types of therapeutic energy, the one or more types of        therapeutic energy comprising one or more of laser radiation,        radio frequency (RF) waves, audio frequency waves, or light from        a light emitting diode (LED), wherein the one or more types of        therapeutic energy have a wavelength of about 200 nm to about        2500 nm and/or frequency of 1-2.4 GHz.-   82. The device of clause 81, wherein the device comprises a laser    configured to generate the laser radiation, the laser comprising one    or more individual fibers configured to conduct the laser radiation,    the laser radiation having a power of about 0.1 to about 5 W per    fiber, wherein the laser radiation is not pulsed.-   83. The device of clause 81, wherein the device comprises an LED    configured to generate light comprising photons configured to be    received by the tissue, the LED arranged on a board with a plurality    of other LEDs configured to generate the same and/or similar light.-   84. The device of clause 81, wherein the device comprises an RF    and/or an audio frequency function generator, the RF and/or audio    frequency comprising a frequency of about 100 Hz-4 MHz.-   85. The device of clause 81, wherein the one or more energy sources    comprise two or more of, or three or more of a laser, an LED, an RF    frequency function generator, and an audio frequency function    generator.-   86. A method for generating one or more types of therapeutic energy    for provision to tissue, the device comprising:    -   generating, with one or more energy sources, one or more types        of therapeutic energy, the one or more types of therapeutic        energy comprising one or more of laser radiation, radio        frequency (RF) waves, audio frequency waves, or light from a        light emitting diode (LED), wherein the one or more types of        therapeutic energy have a wavelength of about 200 nm to about        2500 nm; and    -   directing the one or more types of therapeutic energy toward the        tissue.-   87. The method of clause 86, wherein the generating comprises    generating laser radiation with a laser, the laser comprising one or    more individual fibers configured to conduct the laser radiation,    the laser radiation having a power of about 0.1-5 W per fiber, and    wherein the laser radiation is not pulsed.-   88. The method of clause 86, wherein the generating comprises    generating light with an LED, the light comprising photons    configured to be received by the tissue, the LED arranged on a board    with a plurality of other LEDs configured to generate the same    and/or similar light.-   89. The method of clause 86, wherein the generating comprises    generating RF and/or audio frequency energy with an RF and/or an    audio frequency function generator, the RF and/or audio frequency    comprising a frequency of about 100 Hz-4 MHz.-   90. The method of clause 86, wherein the generating comprises    generating two or more, or three or more, of the types of    therapeutic energy and directing the two or more, or three or more,    types of therapeutic energy toward the tissue.-   91. A device for imaging tissue, comprising:    -   one or more light emitting diodes (LED) configured to emit light        for illuminating the tissue, the light have a wavelength of        about 790-860 nm (IR), and a power of about 0.5-10 W per LED,        and    -   a camera configured to obtain images of the illuminated tissue;    -   wherein the one or more LEDs and the camera are configured to be        moved back and forth over the tissue such that the camera        acquires a plurality of images during the movement.-   92. The device of clause 91, wherein the one or more LEDs are    configured to emit light in the green wavelength range of the    visible spectrum or the infrared wavelength range.-   93. The device of clause 91, further comprising one or more    processors configured to analyze images obtained by the camera by    combining individual images into a common image, and identifying    anatomical features in the tissue based on the common image.-   94. The device of clause 91, further comprising a handle or a stand    coupled to the one or more LEDs and the camera, the handle or the    stand configured to be manipulated by a user such that the light    from the one or more LEDs is directed at the tissue.-   95. The device of clause 91, further comprising a display screen and    a keyboard configured to facilitate entry or selection of    information configured to control the imaging device.-   96. The device of clause 91, wherein one or more veins in tissues    are located using the plurality of images.-   97. The device of clause 91, wherein reflected light from the tissue    is captured by the camera.-   98. The device of clause 91, wherein the one or more processors are    formed by a raspberry pi computing module.

We claim:
 1. A composition with wound healing and tissue regenerativeproperties, the composition comprising two or more of, or three or moreof: a. collagen b. hyaluronic acid c. fucose, d. copper, and e. iron. 2.The composition of claim 1, wherein the collagen comprises collagenfibers.
 3. The composition of claim 1, wherein the collagen comprisespartially or completely hydrolyzed collagen.
 4. The composition of anyone of claim 1, wherein the collagen is acid treated collagen.
 5. Thecomposition of any one of claim 1, wherein the copper comprises a coppersalt.
 6. The composition of claim 5, wherein the copper salt comprisesCuCl₂ or CuCl₃.
 7. The composition of any one of claim 1, wherein theiron comprises an iron salt.
 8. The composition of claim 7, wherein theiron salt comprises FeCl₃.
 9. The composition of any one of claim 1,wherein the composition has a pH in a range of 1 to 4, or 2 to
 3. 10.The composition of any one of claim 1, wherein the composition comprisesa water content in a range of 0% to 20%, or 0% to 10%, or 0% to 5%(wt/wt).
 11. A system configured to be used to treat and/or generatetissue, the system comprising: a device configured to generate one ormore types of therapeutic energy for provision to the tissue, the one ormore types of therapeutic energy comprising one or more of laserradiation, radio frequency (RF) waves, audio frequency waves, or lightfrom a light emitting diode (LED); and a composition with wound healingand/or tissue regenerative properties, the composition comprising two ormore of, or three or more of: a. collagen b. hyaluronic acid c. fucose,d. copper, and e. iron.
 12. The system of claim 11, wherein the collagenis present in the composition in a range of 0.01-0.5% (wt/wt), thehyaluronic acid is present in the composition in a range of 0.1-1.5%(wt/wt), the copper is present in the composition in a range of0.01%-0.1% (wt/wt) and the iron is present in the composition in a rangeof 0.01%-0.1% (wt/wt).
 13. The system of claim 11, wherein the devicecomprises a laser configured to generate the laser radiation, the lasercomprising one or more individual fibers configured to conduct the laserradiation, the laser radiation having a power of about 0.1-5 W perfiber.
 14. The system of claim 11, wherein the device comprises an LEDconfigured to generate light comprising photons configured to bereceived by the tissue, the LED arranged on a board with a plurality ofother LEDs configured to generate the same and/or similar light.
 15. Thesystem of claim 11, wherein the device comprises an RF and/or an audiofrequency function generator, the RF and/or audio frequency comprising afrequency of about 100 Hz-4 MHz.
 16. A device for imaging tissue,comprising: one or more light emitting diodes (LED) configured to emitlight for illuminating the tissue, the light have a wavelength of about790-860 nm (IR), and a power of about 0.5-10 W per LED, and a cameraconfigured to obtain images of the illuminated tissue; wherein the oneor more LEDs and the camera are configured to be moved back and forthover the tissue such that the camera acquires a plurality of imagesduring the movement.
 17. The device of claim 16, wherein the one or moreLEDs are configured to emit light in the green wavelength range of thevisible spectrum or the infrared wavelength range.
 18. The device ofclaim 16, further comprising one or more processors configured toanalyze images obtained by the camera by combining individual imagesinto a common image, and identifying anatomical features in the tissuebased on the common image.
 19. The device of claim 16, furthercomprising a handle or a stand coupled to the one or more LEDs and thecamera, the handle or the stand configured to be manipulated by a usersuch that the light from the one or more LEDs is directed at the tissue.20. The device of claim 18, wherein the one or more processors areformed by a raspberry pi computing module.